IL-8 receptor antagonists

ABSTRACT

This invention relates to novel compounds of Formula (I), and compositions thereof, useful in the treatment of disease states mediated by the chemokine, Interleukin-8 (IL-8).

This application is a 371 of PCT/US01/09216 filed Mar. 23, 2001.

FIELD OF THE INVENTION

This invention relates to novel sulfonamide substituted diphenylthiourea compounds, pharmaceutical compositions, processes for theirpreparation, and use thereof in treating IL-8, GROα, GROβ, GROγ, NAP-2,and ENA-78 mediated diseases.

BACKGROUND OF THE INVENTION

Many different names have been applied to Interleukin-8 (IL8), such asneutrophil attractant/activation protein-1 (NAP-1), monocyte derivedneutrophil chemotactic factor (MDNCF), neutrophil activating factor(NAF), and T-cell lymphocyte chemotactic factor. Interleukin-8 is achemoattractant for neutrophils, basophils, and a subset of T-cells. Itis produced by a majority of nucleated cells including macrophages,fibroblasts, endothelial and epithelial cells exposed to TNF, IL-1α,IL-1β or LPS, and by neutrophils themselves when exposed to LPS orchemotactic factors such as FMLP. M. Baggiolini et al., J. Clin. Invest.84, 1045 (1989); J. Schroder et al, J. Immunol. 139, 3474 (1987) and J.Immunol. 144, 2223 (1990); Strieter, et al., Science 243, 1467 (1989)and J. Biol. Chem. 264, 10621 (1989); Cassatella et al., J. Immunol.148, 3216 (1992).

GROα, GROβ, GROγ and NAP-2 also belong to the chemokine family. LikeIL-8 these chemokines have also been referred to by different names. Forinstance GROα, β, γ have been referred to as MGSAα, β and γ respectively(Melanoma Growth Stimulating Activity), see Richmond et al., J. CellPhysiology 129, 375 (1986) and Chang et al., J. Immunol 148, 451 (1992).All of the chemokines of the α-family which possess the ELR motifdirectly preceding the CXC motif bind to the IL-8 B receptor (CXCR2).

IL-8, GROα, GROβ, GROγ, NAP-2, and ENA-78 stimulate a number of afunctions in vitro. They have all been shown to have chemoattractantproperties for neutrophils, while IL-8 and GROα have demonstratedT-lymphocytes, and basophilic chemotactic activity. In addition IL-8 caninduce histamine release from basophils from both normal and atopicindividuals. GRO-α and IL-8 can in addition, induce lysozomal enzymerelease and respiratory burst from neutrophils. IL-8 has also been shownto increase the surface expression of Mac-1 (CD11b/CD18) on neutrophilswithout de novo protein synthesis. This may contribute to increasedadhesion of the neutrophils to vascular endothelial cells. Many knowndiseases are characterized by massive neutrophil infiltration. As IL-8,GROα, GROβ, GROγ and NAP-2 promote the accumulation and activation ofneutrophils, these chemokines have been implicated in a wide range ofacute and chronic inflammatory disorders including psoriasis andrheumatoid arthritis, Baggiolini et al., FEBS Lett. 307, 97 (1992);Miller et al., Crit. Rev. Immunol. 12, 17 (1992); Oppenheim et al.,Annu. Rev. Immunol. 9, 617 (1991); Seitz et al., J. Clin. Invest. 87,463 (1991); Miller et al., Am. Rev. Respir. Dis. 146, 427 (1992);Donnely et al., Lancet 341, 643 (1993). In addition the ELR chemokines(those containing the amino acids ELR motif just prior to the CXC motif)have also been implicated in angiostasis, Strieter et al., Science 258,1798 (1992).

In vitro, IL-8, GROα, GROβ, GROγ and NAP-2 induce neutrophil shapechange, chemotaxis, granule release, and respiratory burst, by bindingto and activating receptors of the seven-transmembrane, G-protein-linkedfamily, in particular by binding to IL-8 receptors, most notably the IL8β receptor (CXCR2). Thomas et al., J. Biol. Chem. 266, 14839 (1991);and Holmes et al., Science 253, 1278 (1991). The development ofnon-peptide small molecule antagonists for members of this receptorfamily has precedent. For a review see R. Freidinger in: Progress inDrug Research, Vol. 40, pp. 33-98, Birkhauser Verlag, Basel 1993. Hence,the IL-8 receptor represents a promising target for the development ofnovel anti-inflammatory agents.

Two high affinity human IL-8 receptors (77% homology) have beencharacterized: IL-8Rα, which binds only IL8 with high affinity, andIL-8Rβ, which has high affinity for IL-8 as well as for GROα, GROβ, GROγand NAP-2. See Holmes et al., supra; Murphy et al., Science 253, 1280(1991); Lee et al., J. Biol. Chem. 267, 16283 (1992); LaRosa et al., J.Biol. Chem. 267, 25402 (1992); and Gayle et al., J. Biol. Chem. 268,7283 (1993).

There remains a need for treatment, in this field, for compounds, whichare capable of binding to the IL-8 α or β receptor. Therefore,conditions associated with an increase in IL-8 production (which isresponsible for chemotaxis of neutrophil and T-cells subsets into theinflammatory site) would benefit by compounds, which are inhibitors ofIL-8 receptor binding.

SUMMARY OF THE INVENTION

This invention provides for a method of treating a chemokine mediateddisease, wherein the chemokine is one which binds to an IL-8 a or breceptor and which method comprises administering an effective amount ofa compound of Formula (I) or a pharmaceutically acceptable salt thereof.In particular the chemokine is IL-8.

This invention also relates to a method of inhibiting the binding ofIL-8 to its receptors in a mammal in need thereof which comprisesadministering to said mammal an effective amount of a compound ofFormula (I).

The present invention also provides for the novel compounds of Formula(I), and pharmaceutical compositions comprising a compound of Formula(I), and a pharmaceutical carrier or diluent.

Compounds of Formula (I) useful in the present invention are representedby the structure:

wherein:

R is selected from the group consisting of cyano, OR₁₁, C(O)NR₁₅R₁₆,R₁₈, C(O)OR₁₁, C(O)R₁₁, and S(O)₂R₁₇;

R_(b) is independently selected from the group consisting of hydrogen,NR₆R₇, OH, OR_(a), C₁₋₅alkyl, aryl, arylC₁₋₄alkyl, aryl C₂₋₄alkenyl;cycloalkyl, cycloalkyl C₁₋₅ alkyl, heteroaryl, heteroarylC₁₋₄alkyl,heteroarylC₂₋₄ alkenyl, heterocyclic, heterocyclic C₁₋₄alkyl, and aheterocyclic C₂₋₄alkenyl moiety, all of which moieties are optionallysubstituted one to three times independently by a substituent selectedfrom the group consisting of halogen, nitro, halosubstituted C₁₋₄ alkyl,C₁₋₄ alkyl, amino, mono and di-C₁₋₄ alkyl substituted amine, OR_(a),C(O)R_(a), NR_(a)C(O)OR_(a), OC(O)NR₆R₇, hydroxy, NR₉C(O)R_(a),S(O)_(t)R_(a), C(O)NR₆R₇, C(O)OH, C(O)OR_(a), S(O)_(t)NR₆R₇, andNHS(O)_(t)R_(a); or the two R_(b) substituents join to form a 3-10membered ring, and containing, in addition to optionally substitutedC₁₋₉ alkyl, independently, 0 to 3 substituents selected from the groupconsisting of NR_(a), C(O), O, S, SO, and SO₂ moieties which areunsaturated or saturated.

R_(a) is selected from the group consisting if alkyl, aryl,arylC₁₋₄alkyl, heteroaryl, heteroaryl C₁₋₄alkyl, heterocyclic, COOR_(a),and a heterocyclic C₁₋₄alkyl moiety, all of which moieties areoptionally substituted;

m is an integer having a value of 1 to 3;

m′ is 0, or an integer having a value of 1 or 2;

n is an integer having a value of 1 to 3;

q is 0, or an integer having a value of 1 to 10;

t is 0, or an integer having a value of 1 or 2;

s is an integer having a value of 1 to 3;

R₁ is independently selected from the group consisting of

hydrogen, halogen, nitro, cyano, C₁₋₁₀ alkyl, halosubstituted C₁₋₁₀alkyl, C₂₋₁₀ alkenyl, C₁₋₁₀ alkoxy, halosubstituted C₁₋₁₀alkoxy,azide,S(O)_(t)R₄, (CR₈R₈)_(q)S(O)_(t)R₄, hydroxy, hydroxy substitutedC₁₋₄alkyl,aryl, aryl C₁₋₄ alkyl, aryl C₂₋₁₀ alkenyl, aryloxy, aryl C₁₋₄alkyloxy, heteroaryl, heteroarylalkyl, heteroaryl C₂₋₁₀ alkenyl,heteroaryl C₁₋₄ alkyloxy, heterocyclic, heterocyclic C₁₋₄alkyl,heterocyclicC₁₋₄alkyloxy, heterocyclicC₂₋₁₀ alkenyl, (CR₈R₈)_(q)NR₄R₅,(CR₈R₈)_(q)C(O)NR₄R₅, C₂₋₁₀ alkenyl C(O)NR₄R₅, (CR₈R₈)_(q)C(O)NR₄R₁₀,S(O)₃R₈, (CR₈R₈)_(q)C(O)R₁₁, C₂₋₁₀ alkenyl C(O)R₁₁,

C₂₋₁₀ alkenyl C(O)OR₁₁, (CR₈R₈)_(q)C(O)OR₁₁, (CR₈R₈)_(q)OC(O)R₁₁,(CR₈R₈)_(q)NR₄C(O)R₁₁, (CR₈R₈)_(q)C(NR₄)NR₄R₅, (CR₈R₈)_(q)NR₄C(NR₅)R₁₁,(CR₈R₈)_(q)NHS(O)₂R₁₃, and (CR₈R₈)_(q)S(O)₂NR₄R₅, or two R₁ moietiestogether form O—(CH₂)_(s)O or a 5 to 6 membered saturated or unsaturatedring, wherein the alkyl, aryl, arylalkyl, heteroaryl, or heterocyclicmoieties are optionally substituted;

R₄ and R₅ are independently selected from the group consisting ofhydrogen, optionally substituted C₁₋₄ alkyl, optionally substitutedaryl, optionally substituted aryl C₁₋₄alkyl, optionally substitutedheteroaryl, optionally substituted heteroaryl C₁₋₄alkyl, heterocyclic,and heterocyclicC₁₋₄alkyl; or R₄ and R₅ together with the nitrogen towhich they are attached form a 5 to 7 member ring which optionallycomprises an additional heteroatom selected from the group consisting ofO, N and S;

R₆ and R₇ are independently selected from the group consisting ofhydrogen, C₁₋₄ alkyl, heteroaryl, aryl, alkyl aryl, and alkyl C₁₋₄heteroalkyl; or R₆ and R₇ together with the nitrogen to which they areattached form a 5 to 7 member ring which ring optionally contains anadditional heteroatom selected from the group consisting of oxygen,nitrogen and sulfur, which ring is optionally substituted;

Y is selected from the group consisting of hydrogen, halogen, nitro,cyano, halosubstituted C₁₋₁₀ alkyl, C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₁₋₁₀alkoxy, halosubstituted C₁₋₁₀ alkoxy, azide, (CR₈R₈)_(q)S(O)_(t)R_(a),(CR₈R₈)_(q)OR_(a), hydroxy, hydroxy substituted C₁₋₄alkyl, aryl, arylC₁₋₄ alkyl, aryloxy, arylC₁₋₄ alkyloxy, aryl C₂₋₁₀ alkenyl, heteroaryl,heteroarylalkyl, heteroaryl C₁₋₄ alkyloxy, heteroaryl C₂₋₁₀ alkenyl,heterocyclic, heterocyclic C₁₋₄alkyl, heterocyclicC₂₋₁₀ alkenyl,(CR₈R₈)_(q)NR₄R₅, C₂₋₁₀ alkenyl C(O)NR₄R₅, (CR₈R₈)_(q)C(O)NR₄R₅,(CR₈R₈)_(q)C(O)NR₄R₁₀, S(O)₃R₈, (CR₈R₈)_(q)C(O)R₁₁, C₂₋₁₀alkenylC(O)R₁₁, (CR₈R₈)_(q)C(O)OR₁₁, C₂₋₁₀alkenylC(O)OR₁₁,(CR₈R₈)_(q)OC(O)R₁₁, (CR₈R₈)_(q)NR₄C(O)R₁₁, (CR₈R₈)_(q)NHS(O)_(t)R₁₃,(CR₈R₈)_(q)S(O)_(t)NR₄R₅, (CR₈R₈)_(q)C(NR₄)NR₄R₅, and(CR₈R₈)_(q)NR₄C(NR₅)R₁₁; or two Y moieties together form O—(CH₂)_(s)—Oor a 5 to 6 membered saturated or unsaturated ring; wherein the alkyl,aryl, arylalkyl, heteroaryl, heteroaryl alkyl, heterocyclic,heterocyclicalkyl groups are optionally substituted;

R₈ is hydrogen or C₁₋₄ alkyl;

R₉ is hydrogen or a C₁₋₄ alkyl;

R₁₀ is C₁₋₁₀ alkyl C(O)₂R₈;

R₁₁ is selected from the group consisting of hydrogen, optionallysubstituted C₁₋₄ alkyl, optionally substituted aryl, optionallysubstituted aryl C₁₋₄alkyl, optionally substituted heteroaryl,optionally substituted heteroarylC₁₋₄alkyl, optionally substitutedheterocyclic, and optionally substituted heterocyclicC₁₋₄alkyl; and

R₁₃ is selected from the group consisting of C₁₋₄ alkyl, aryl, arylC₁₋₄alkyl, heteroaryl, heteroarylC₁₋₄alkyl, heterocyclic, andheterocyclicC₁₋₄alkyl;

or a pharmaceutically acceptable salt thereof.

DETAILED DESCRIPTION OF THE INVENTION

The compounds of Formula (I), may also be used in association with theveterinary treatment of mammals, other than humans, in need ofinhibition of IL-8 or other chemokines which bind to the IL-8 α and βreceptors. Chemokine mediated diseases for treatment, therapeutically orprophylactically, in animals include disease states such as those notedherein in the Methods of Treatment section.

Suitably, R_(b) is independently hydrogen, NR₆R₇, OH, OR_(a), C₁₋₄alkyl,aryl, arylC₁₋₄alkyl, aryl C₂₋₄alkenyl, heteroaryl, heteroarylC₁₋₄alkyl,heteroarylC₂₋₄ alkenyl, heterocyclic, heterocyclic C₁₋₄alkyl, or aheterocyclic C₂₋₄alkenyl moiety, all of which moieties may be optionallysubstituted one to three times independently by halogen; nitro;halosubstituted C₁₋₄alkyl; C₁₋₄ alkyl; amino, mono or di-C₁₋₄ alkylsubstituted amine; cycloalkyl, cycloalkyl C₁₋₅ alkyl, OR_(a); C(O)R_(a);NR_(a)C(O)OR_(a); OC(O)NR₆R₇; aryloxy; aryl C₁₋₄ oxy; hydroxy; C₁₋₄alkoxy; NR₉C(O)R_(a); S(O)_(m′)R_(a); C(O)NR₆R₇; C(O)OH; C(O)OR_(a);S(O)_(t)NR₆R₇; NHS(O)_(t)R_(a); Alternatively, the two R_(b)substituents can join to form a 3-10 membered ring, optionallysubstituted and containing, in addition to carbon, independently, 1 to 3NR₉, O, S, SO, or SO₂ moities which can be optionally substituted.

Suitably, R_(a) is an alkyl, aryl, arylC₁₋₄alkyl, heteroaryl, heteroarylC₁₋₄alkyl, heterocyclic, or a heterocyclic C₁₋₄alkyl moiety, all ofwhich moieties may be optionally substituted.

Suitably R₁ is independently selected from hydrogen; halogen; nitro;cyano; halosubstituted C₁₋₁₀ alkyl, such as CF₃; C₁₋₁₀ alkyl, such asmethyl, ethyl, isopropyl, or n-propyl; C₂₋₁₀ alkenyl; C₁₋₁₀ alkoxy, suchas methoxy, or ethoxy; halosubstituted C₁₋₁₀ alkoxy, such astrifluoromethoxy; azide; (CR₈R₈)_(q)S(O)_(t)R₄, wherein t is 0, 1 or 2;hydroxy; hydroxy C₁₋₄alkyl, such as methanol or ethanol; aryl, such asphenyl or naphthyl; aryl C₁₋₄ alkyl, such as benzyl; aryloxy, such asphenoxy; aryl C₁₋₄ alkyloxy, such as benzyloxy; heteroaryl;heteroarylalkyl; heteroaryl C₁₋₄ alkyloxy; aryl C₂₋₁₀ alkenyl;heteroaryl C₂₋₁₀ alkenyl; heterocyclic C₂₋₁₀ alkenyl; (CR₈R₈)_(q)NR₄R₅;C₂₋₁₀ alkenyl C(O)NR₄R₅; (CR₈R₈)_(q)C(O)NR₄R₅; (CR₈R₈)_(q)C(O)NR₄R₁₀;S(O)₃H; S(O)₃R₈; (CR₈R₈)_(q)C(O)R₁₁; C₂₋₁₀ alkenyl C(O)R₁₁; C₂₋₁₀alkenyl C(O)OR₁₁; (CR₈R₈)_(q)C(O)R₁₁; (CR₈R₈)_(q)C(O)OR₁₁;(CR₈R₈)_(q)OC(O)R₁₁; (CR₈R₈)_(q)NR₄C(O)R₁₁; (CR₈R₈)_(q)C(NR₄)NR₄R₅;(CR₈R₈)_(q)NR₄C(NR₅)R₁₁; (CR₈R₈)_(q)NHS(O)_(t)R₁₃;(CR₈R₈)_(q)S(O)_(t)NR₄R₅. All of the aryl, heteroaryl, and heterocycliccontaining moieties may be optionally substituted as defined hereinbelow.

For use herein the term “the aryl, heteroaryl, and heterocycliccontaining moieties” refers to both the ring and the alkyl, or ifincluded, the alkenyl rings, such as aryl, arylalkyl, and aryl alkenylrings. The term “moieties” and “rings” may be interchangeably usedthroughout.

Suitably, R₄ and R₅ are independently hydrogen, optionally substitutedC₁₋₄ alkyl, optionally substituted aryl, optionally substituted arylC₁₋₄alkyl, optionally substituted heteroaryl, optionally substitutedheteroaryl C₁₋₄alkyl, heterocyclic, heterocyclicC₁₋₄ alkyl, or R₄ and R₅together with the nitrogen to which they are attached form a 5 to 7member ring which may optionally comprise an additional heteroatomselected from O/N/S.

Suitably, R₈ is independently hydrogen or C₁₋₄ alkyl.

Suitably, R₉ is hydrogen or a C₁₋₄ alkyl;

Suitably, q is 0 or an integer having a value of 1 to 10.

Suitably, R₁₀ is C₁₋₁₀ alkyl C(O)₂R₈, such as CH₂C(O)₂H or CH₂C(O)₂CH₃.

Suitably, R₁₁ is hydrogen, C₁₋₄ alkyl, aryl, aryl C₁₋₄ alkyl,heteroaryl, heteroaryl C₁₋₄alkyl, heterocyclic, or heterocyclicC₁₋₄alkyl.

Suitably, R₁₂ is hydrogen, C₁₋₁₀ alkyl, optionally substituted aryl oroptionally substituted arylalkyl.

Suitably, R₁₃ is C₁₋₄alkyl, aryl, arylalkyl, heteroaryl,heteroarylC₁₋₄alkyl, heterocyclic, or heterocyclicC₁₋₄alkyl, wherein allof the aryl, heteroaryl and heterocyclic containing moieties may all beoptionally substituted.

Suitably, Y is independently selected from hydrogen; halogen; nitro;cyano; halosubstituted C₁₋₁₀ alkyl; C₁₋₁₀ alkyl; C₂₋₁₀ alkenyl; C₁₋₁₀alkoxy; halosubstituted C₁₋₁₀ alkoxy; azide; (CR₈R₈)_(q)S(O)_(t)R_(a);hydroxy; hydroxyC₁₋₄alkyl; aryl; aryl C₁₋₄ alkyl; aryloxy; arylC₁₋₄alkyloxy; heteroaryl; heteroarylalkyl; heteroaryl C₁₋₄ alkyloxy;heterocyclic, heterocyclic C₁₋₄alkyl; aryl C₂₋₁₀ alkenyl; heteroarylC₂₋₁₀ alkenyl; heterocyclic C₂₋₁₀ alkenyl; (CR₈R₈)_(q)NR₄R₅; C₂₋₁₀alkenyl C(O)NR₄R₅; (CR₈R₈)_(q)C(O)NR₄R₅; (CR₈R₈)_(q)C(O)NR₄R₁₀; S(O)₃H;S(O)₃R₈; (CR₈R₈)_(q)C(O)R₁₁; C₂₋₁₀ alkenyl C(O)R₁₁; C₂₋₁₀ alkenylC(O)OR₁₁; (CR₈R₈)_(q)C(O)OR₁₂; (CR₈R₈)_(q)OC(O)R₁₁;(CR₈R₈)_(q)C(NR₄)NR₄R₅; (CR₈R₈)_(q)NR₄C(NR₅)R₁₁; (CR₈R₈)_(q)NR₄C(O)R₁₁;(CR₈R₈)_(q)NHS(O)_(t)R₁₃; or (CR₈R₈)_(q)S(O)_(t)NR₄R₅; or two Y moietiestogether may form O—(CH₂)_(s)—O or a 5 to 6 membered saturated orunsaturated ring. The aryl, heteroaryl and heterocyclic containingmoieties noted above may all be optionally substituted as definedherein.

Suitably s is an integer having a value of 1 to 3.

When Y forms a dioxybridge, s is preferably 1. When Y forms anadditional unsaturated ring, it is preferably 6 membered resulting in anaphthylene ring system. These ring systems may be substituted 1 to 3times by other Y moieties as defined above.

Suitably, R_(a) is an alkyl, aryl C₁₋₄ alkyl, heteroaryl,heteroaryl-C₁₋₄alkyl, heterocyclic, or a heterocyclicC₁₋₄ alkyl, whereinall of these moieties may all be optionally substituted.

Y is preferably a halogen, C₁₋₄ alkoxy, optionally substituted aryl,optionally substituted aryloxy or arylalkoxy, methylene dioxy, NR₄R₅,thio C₁₋₄alkyl, thioaryl, halosubstituted alkoxy, optionally substitutedC₁₋₄ alkyl, or hydroxy alkyl. Y is more preferably mono-substitutedhalogen, disubstituted halogen, mono-substituted alkoxy, disubstitutedalkoxy, methylenedioxy, aryl, or alkyl, more preferably these groups aremono or di-substituted in the 2′-position or 2′-, 3′-position.

While Y may be substituted in any of the ring positions, n is preferablyone. While both R₁ and Y can both be hydrogen, it is preferred that atleast one of the rings is substituted, preferably both rings aresubstituted.

As used herein, “optionally substituted” unless specifically definedshall mean such groups as halogen, such as fluorine; chlorine, bromineor iodine; hydroxy; hydroxy substituted C₁₋₁₀alkyl; (C₈R₈)_(q)OR₄;C₁₋₁₀alkoxy, such as methoxy or ethoxy; two substituents together mayform O—(CH₂)_(s)—O; S(O)_(m′)C₁₋₁₀ alkyl, wherein m′ is 0, 1 or 2, suchas methyl thio, methyl sulfinyl or methyl sulfonyl; amino, mono &di-substituted amino, such as in the NR₄R₅ group; NHC(O)R₄; C(O)NR₄R₅;C(O)OR₄; S(O)_(t)NR₄R₅; NHS(O)_(t)R₂₀, C₁₋₁₀ alkyl, such as methyl,ethyl, propyl, isopropyl, or t-butyl; halosubstituted C₁₋₁₀ alkyl, suchCF₃; an optionally substituted aryl, such as phenyl, or an optionallysubstituted arylalkyl, such as benzyl or phenethyl, optionallysubstituted heterocylic, optionally substituted heterocyclicalkyl,optionally substituted heteroaryl, optionally substituted heteroarylalkyl, wherein these aryl , heteroaryl, or heterocyclic moieties may besubstituted one to two times by halogen; hydroxy; hydroxy substitutedalkyl; C₁₋₁₀ alkoxy; S(O)_(m′)C₁₋₁₀ alkyl; amino, mono & di-substitutedalkyl amino, such as in the NR₄R₅ group; C₁₋₁₀ alkyl, or halosubstitutedC₁₋₁₀ alkyl, such as CF₃.

R₂₀ is suitably C₁₋₄ alkyl, aryl, aryl C₁₋₄alkyl, heteroaryl,heteroarylC₁₋₄alkyl, heterocyclic, or heterocyclicC₁₋₄alkyl.

Suitable pharmaceutically acceptable salts are well known to thoseskilled in the art and include basic salts of inorganic and organicacids, such as hydrochloric acid, hydrobromic acid, sulphuric acid,phosphoric acid, methane sulphonic acid, ethane sulphonic acid, aceticacid, malic acid, tartaric acid, citric acid, lactic acid, oxalic acid,succinic acid, fumaric acid, maleic acid, benzoic acid, salicylic acid,phenylacetic acid and mandelic acid. In addition, pharmaceuticallyacceptable salts of compounds of Formula (I) may also be formed with apharmaceutically acceptable cation. Suitable pharmaceutically acceptablecations are well known to those skilled in the art and include alkaline,alkaline earth, ammonium and quaternary ammonium cations.

The following terms, as used herein, refer to:

“halo”—all halogens, that is chloro, fluoro, bromo and iodo.

“C₁₋₁₀alkyl” or “alkyl”—both straight and branched chain moieties of 1to 10 carbon atoms, unless the chain length is otherwise limited,including, but not limited to, methyl, ethyl, n-propyl, iso-propyl,n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl and the like.

“cycloalkyl” is used herein to mean cyclic moiety, preferably of 3 to 8carbons, including but not limited to cyclopropyl, cyclopentyl,cyclohexyl, and the like.

“alkenyl” is used herein at all occurrences to mean straight or branchedchain moiety of 2-10 carbon atoms, unless the chain length is limitedthereto, including, but not limited to ethenyl, 1-propenyl, 2-propenyl,2-methyl-1-propenyl, 1-butenyl, 2-butenyl and the like.

“aryl”—phenyl and naphthyl;

“heteroaryl” (on its own or in any combination, such as “heteroaryloxy”,or “heteroaryl alkyl”)—a 5-10 membered aromatic ring system in which oneor more rings contain one or more heteroatoms selected from the groupconsisting of N, O or S, such as, but not limited, to pyrrole, pyrazole,furan, thiophene, quinoline, isoquinoline, quinazolinyl, pyridine,pyrimidine, oxazole, tetrazole, thiazole, thiadiazole, triazole,imidazole, or benzimidazole.

“heterocyclic” (on its own or in any combination, such as“heterocyclicalkyl”)—a saturated or partially unsaturated 4-10 memberedring system in which one or more rings contain one or more heteroatomsselected from the group consisting of N, O, or S; such as, but notlimited to, pyrrolidine, piperidine, piperazine, morpholine,tetrahydropyran, thiomorpholine, or imidazolidine. Furthermore, sulfurmay be optionally oxidized to the sulfone or the sulfoxide.

“arylalkyl” or “heteroarylalkyl” or “heterocyclicalkyl” is used hereinto mean C₁₋₁₀ alkyl, as defined above, attached to an aryl, heteroarylor heterocyclic moiety, as also defined herein, unless otherwiseindicated.

“sulfinyl”—the oxide S(O) of the corresponding sulfide, the term “thio”refers to the sulfide, and the term “sulfonyl” refers to the fullyoxidized S(O)₂ moiety.

“wherein two R₁ moieties (or two Y moieties) may together form a 5 or 6membered saturated or unsaturated ring” is used herein to mean theformation of an aromatic ring system, such as naphthalene, or is aphenyl moiety having attached a 6 membered partially saturated orunsaturated ring such as a C₆ cycloalkenyl, i.e. hexene, or a C₅cycloalkenyl moiety, such as cyclopentene.

Illustrative compounds of Formula (I) include:

N-(2-bromophenyl)-N′-[4-chloro-2-hydroxy-3-(N″,N″dimethylaminosulfonyl)phenyl]cyanoguanidine;

N-[4chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2,3-dichlorophenyl)cyanoguanidine;

N-(2-bromophenyl)-N′-[4-chloro2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]cyanoguanidine;

N-(2,3-dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]cyanoguanidine;

N-phenyl-N′-[4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl[aminosulfonylphenyl]cyanoguanidine;

N-(2-bromophenyl)-N′-[4chloro-2-hydroxy-3-[R-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]cyanoguanidine;

N-(2,3-dichlorophenyl)-N′-[4chloro-2-hydroxy-3-[R-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]cyanoguanidine;

N-(2-bromophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-isoxazolidinylaminosulfonylphenyl]cyanoguanidine;

N-(2,3-dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-isoxazolidinylaminosulfonylphenyl]cyanoguanidine;

N-(2-bromophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-tetrahydroisoxazylaminosulfonyl)phenyl]cyanoguanidine;

N-(2,3-dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-tetrahydroisoxazylaminosulfonyl)phenyl]cyanoguanidine;

N-(2,3-dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-(4-thiomorpholinylaminosulfonyl)phenyl]cyanoguanidine;

N-[4-chloro-2-hydroxy-3-[N″,N″-dimethylaminosulfonyl]phenyl]-N′-(2-bromophenyl)propylguanidine;

N-(2-bromophenyl)-N′-[4-chloro-2-hydroxy-3-(4-oxidothiomorpholino)aminosulfonylphenyl]cyanoguanidine;

N-(2,3-chlorophenyl)-N′-[4-chloro-2-hydroxy-3-(4-oxidothiomorpholino)aminosulfonylphenyl]cyanoguanidine;

N-(2-bromophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-methylpiperazino)aminosulfonylphenyl]cyanoguanidine;

N-(2,3-dichlorophenyl)-N′-[4-chloro-2-hydroxy-3(N″-methylpiperazino)aminosulfonylphenyl]cyanoguanidine;

N-(2-bromophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-ethylmorpholino)aminosulfonylphenyl]cyanoguanidine;

N-(2,3-dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-ethylmorpholino)aminosulfonylphenyl]cyanoguanidine;

N-(2-bromophenyl)-N′-{4-chloro-2-hydroxy-3-[N″-ethyl-2-(2-ethylpyrrolidino)]aminosulfonylpheny}cyanoguanidine;

N-(2,3dichlorophenyl)-N′-{4-chloro-2-hydroxy-3-[N″-ethyl-2-(2-ethylpyrrolidino)]aminosulfonylpheny)cyanoguanidine;

N-(2-bromophenyl)-N′-{4-chloro-2-hydroxy-3-[S-(+)-(2-carboxy)pyrrolidin-1-yl]aminosulfonylpheny}cyanoguanidine;

N-(2,3-dichlorophenyl)-N′-(4-chloro-2-hydroxy-3-[S-(+)-(2-carboxy)pyrrolidin-1-yl]aminosulfonylpheny}cyanoguanidine;

N-(2-bromo-3-fluorophenyl)-N′-[4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]sulfonylphenyl]cyanoguanidine;

N-(2-phenoxyphenyl)-N′-[4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]sulfonylphenyl]cyanoguanidine;and

N-(2-benzoxyphenyl)-N′-[4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]sulfonylphenyl]cyanoguanidine;

or a pharmaceutically acceptable salt thereof.

METHODS OF PREPARATION

The compounds of Formulas (I) may be obtained by applying syntheticprocedures, some of which are illustrated in the Schemes below.

Scheme 1

a) i) NCS, AcOH, H₂O, ii) LiOH, MeOH b) H₂SO₄, HNO₃ c) KOH, MeOH d)PCl₅,POCl₃ e) NR′R″H, Et₃N f) NaH, H₂O g) Pd/C, H₂ h)RCNO, DMF i) TBSCl,Imidazole j) MsCl, Et₃N k) NH₂CN, Hunig's base 1) CsF/TBAF, MeOH/THF

The desired 4-chloro-N-(3-sulfonamido-2-hydroxyphenyl)-N″-phenylcyanoguanidine can be synthesized from the commercially available2,6-dichloro thiophenol using procedure elaborated in scheme 1. Thethiol can be oxidized to the sulfonyl halide using a halogenating agentsuch as NCS, NBS, chlorine or bromine in the presence of a proticsolvent such as alcohol, acetic acid or water. The sulfonyl halide canbe hydrolyzed by using a metal hydroxide such as lithium or potassiumhydroxide to form the corresponding sulfonic acid salt.

The sulfonic acid salt can then be nitrated under nitration conditionssuch as nitric acid in a solvent of strong acid such as sulfuric acid toform the nitro phenyl sulfonic acid 3-scheme 1. The sulfonic acid3-scheme 1 can be converted to the sulfonamide 5-scheme 1 using a threestep procedure involving the formation of the metal salt using a basesuch as potassium hydroxide, sodium hydride or sodium carbonate to form4-scheme 1. The sulfonic acid salt is then converted to the sulfonylchloride using PCl₅ with POCl₃ as a solvent. The sulfonyl chloride canthen be converted to the corresponding sulfonamide using the desiredamine HNR′R″ in triethyl amine at temperatures ranging from −78° C. to25° C. to form the corresponding sulfonamide 5-scheme 1.

The chlorine ortho to the nitro group can be selectively hydrolyzedusing sodium hydride/water in THF at room temperature to form thephenol. The nitro can be reduced by conditions well known in the artsuch as hydrogen and palladium on carbon to form the correspondinganiline 6scheme 1. The aniline can then be coupled with a commerciallyavailable thioisocyanate to form the desired thiourea.

The phenol in thiourea can be protected with TBSCl to form thecorresponding compounds 7-scheme 1. The protected thiourea can beconverted to the corresponding carbodiimide 8-scheme 1 usingmethanesulfonyl chloride and triethyl amine at 0° C. The carbodiimde canthen be converted to the corresponding protected cyanoguanidine usingcyanamide and Hunig's base, followed by desilylation with CsF or TBAF toform the desired the cyanoguanidine 9-scheme1.

If the desired hydroxyaniline 6-Scheme 1 is not commercially available,it can be prepared as outlined in Scheme 2: Commercially availablesubstituted 3-chloroanilines 1-scheme-2 can be converted to the amide2-scheme-2 using standard conditions well known in the art such aspivavolyl chloride and triethylamine in a suitable organic solvent suchas methylene chloride. The amide 2-scheme-2 can be converted to thebenzoxazole 3-scheme-2 using an excess amount of a strong base such asbutyllithium in a suitable organic solvent such as THF under reducedreaction temperatures (−20 to −40° C.) followed by quenching thereaction with sulfur dioxide gas and converting resulting sulfinic acidsalt to the sulfonyl chloride 3-scheme-2 using standard conditions wellknown in the art such as sulfuryl chloride in a suitable organic solventsuch as methylene chloride. The sulfonyl chloride 3-scheme-2 can betransformed to the sulfonamide 4-scheme-2 using standard conditions wellknown in the art by reacting it with the amine HN(R_(b))₂ in thepresence of a suitable amine base such as triethylamine in a suitableorganic solvent such as methylene chloride. The desired phenolaniline 5can be obtained from the benzoxazole 4scheme-2 using standard hydrolysisconditions well known in the art such as suilfric acid in water andheating at 85° C.

SYNTHETIC EXAMPLES

The invention will now be described by reference to the followingexamples, which are merely illustrative and are not to be construed as alimitation of the scope of the present invention. All temperatures aregiven in degrees centigrade, all solvents are highest available purityand all reactions run under anhydrous conditions in an argon atmosphereunless otherwise indicated.

In the Examples, all temperatures are in degrees Centigrade (°C.). Massspectra were performed upon a VG Zab mass spectrometer using fast atombombardment, unless otherwise indicated. ¹H-NMR (hereinafter “NMR”)spectra were recorded at 250 MHz using a Bruker AM 250 or Am 400spectrometer. Multiplicities indicated are: s=singlet, d=doublet,t=triplet, q=quartet, m=multiplet and br indicates a broad signal. Sat.indicates a saturated solution, eq indicates the proportion of a molarequivalent of reagent relative to the principal reactant.

Examples 1 & 2 Preparation ofN-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2,3-dichlorophenyl)cyanoguanidineandN-(2-Bromophenyl)-N′-[4-chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]cyanoguanidine2,6-Dichloro-3-nitrobenzenesulfonic Acid

Lithium hydroxide hydrate (8.96 g, 0.214 mol) was added to a solution of2,6-dichlorobenzenesulfonyl chloride (35 g, 0.146 mol) in MeOH (300 mL)and the reaction was allowed to stir at room temperature for 16 hr. Thereaction mixture was filtered to remove suspended solids and thenconcentrated. The resulting solid was dried in vacuo overnight to removeany residual MeOH. The solid was then dissolved in H₂SO₄ (300 mL) andchilled in an ice bath. A solution of H₂SO₄ (35 mL) and HNO₃ (70%, 10.7mL) was slowly added to the above reaction over 90 min. The reaction wasallowed to warm up to room temperature overnight and then slowly pouredinto ice water (1200 mL) and extracted with EtOAc. The combined organiclayers were dried (MgSO₄) and concentrated to yield2,6-dichloro-3-nitrobenzenesulfonic acid (37.38 g, 96%) as thedihydrate. EI-MS (m/z) 270 (M⁻).

2,6-Dichloro-3-nitrobenzenesulfonyl Chloride

Potassium hydroxide (11.54 g, 0.206 mol) was added to a solution of2,6-dichloro-3-nitrobenzenesulfonic acid dihydrate (37.38 g, 0.137 mol)in MeOH (500 mL) and the reaction was allowed to stir at roomtemperature for 14 hr. The reaction mixture was concentrated and theresulting solid was dried in vacuo overnight. To this was added PCl₅(29.60 g, 0.142 mol) followed by POCl₃ (400 mL) and the mixture wasrefluxed overnight. The reaction was then cooled to room temperature andconcentrated. The resulting mixture was taken up in EtOAc and chilled inan ice bath. Ice chunks were slowly added to the reaction mixture toquench any leftover PCl₅. When bubbling ceased, water was added and thereaction mix was extracted with EtOAc. The organic layer was dried(MgSO₄) and concentrated to yield 2,6-Dichloro-3-nitrobenzenesulfonylchloride (31.4 g, 79%).

¹H NMR (DMSO-d₆) δ 7.88 (d, 1H), 7.75 (d, 1H).

The following is the general procedure for sulfonamide formation:

N,N-Dimethyl-2,6-dichloro-3-nitrobenzenesulfonamide

Into a solution of 2,6-dichloro-3-nitrobenzenesulfonyl chloride (200 mg,0.69 mmol) in 15 mL of dichloromethane at −78° C. was added dropwise asolution of dimethylamine (2.0 M in MeOH, 0.345 mL, 0.69 mmol) andtriethylamine (0.14 mL, 1.04 mmol) in 10 mL of dichloromethane. Themixture was warmed to room temperature and stirred for 16 hours. Themixture was acidified to pH>1 with 1N aq. HCl, then extracted with ethylacetate. The combined organic layer was then concentrated to give thecrude material. Column chromatography on silica gel, eluting with ethylacetate/hexane (30/70, v/v/), gave the desired (240 mg, 70%). EI-MS(m/z) 298 (M⁻).

The following is the general procedure for the hydrolysis ofdichlorosulfonamide to phenol:

N,N-Dimethyl-6chloro-2-hydroxy-3-nitrobenzenesulfonamide

A mixture of N,N-dimethyl-2,6-dichloro-3-nitrobenzenesulfonamide (2.64g, 8.83 mmol), 60% sodium hydride (1.06 g, 26.5 mmol) and water (191 mg,10.6 mmol) was heated to 35° C. while kept under argon atmosphere for 16hours. The solvent was evaporated. when The reaction was almost completeas indicated by ¹H NMR. The residue was diluted with ethyl acetate andwashed with 1N aq. HCl. The solvent was concentrated to give the crudematerial. Column chromatography on silica gel, eluting with ethylacetate/hexane/acetic acid (40/58/2, v/v/v), gave the desired product(2.3 g, 93%). EI-MS (m/z)279.5 (M⁻).

The following is the general procedure for the hydrogenation of nitrocompound to aniline:

N,N-Dimethyl-3-amino-6-chloro-2-hydroxybenzenesulfonamide

To a solution ofN,N-dimethyl-6-chloro-2-hydroxy-3-nitrobenzenesulfonamide (2.3 g, 8.2mmol) in ethyl acetate, was added 10% Pd/C (2.0 g). The mixture wasflushed with argon, and then stirred under a hydrogen atmosphere atballoon pressure for 3 hours at room temperature. The mixture wasfiltered through celite and the celite was washed with methanol. Thesolvent was evaporated to give the desired product (2.0 g, 97%). EI-MS(m/z) 249.5 (M⁻).

Preparation of N-(3,4-Dichlorophenyl)2,2-dimethyl-propionamide

3,4-dichloroaniline (150 g) in TBME (1 L) was cooled to 10-15° C. 30% aqNaOH (141 g, 1.14 equiv) was added, and the solution stirred vigorouslyvia overhead mechanical stirrer. Trimethylacetyl chloride (“PivCl”, 126mL) was added at such a rate as to keep the internal temperature below30° C. During this addition, the solution mixture becomes thick withwhite solid product. When the addition was complete (10-15 min), themixture was heated to 30-35° C. for 1 hr, and then allowed to cool. Thereaction mixture was held at −5° C. (overnight), and then filtered,rinsing first with 90:10 water/MeOH (600 mL) and then water (900 mL).Drying under vacuum yielded 195 g (86%) product, as off-white crystals.LCMS m/z 246(M−H)⁺.

Preparation of 2-tert-Butyl-6-chloro-benzooxazole-7-sulfonyl Chloride

The solution of N-(3,4dichloro-phenyl)2,2-dimethyl-propionamide (10 g,41 mmol) in dry THF (100 mL) was cooled to −72° C. under argon. n-Butyllithium (1.6M in hexane, 64 mL, 102 mmol) was added dropwise. Thesolution warmed to ca −50° C. over 45 minutes, and then was kept in the−25-−10° C. range for 2 hrs. The solution was then recooled to −78° C.,and sulfur dioxide was bubbled through the solution for 30 min. Thesolution was then allowed to warm to room temperature for 2 h, and a Arstream was bubbled through the solution, with a gas outlet provided sothat any excess sulfur dioxide could escape during the warming. The THFsolution was cooled in an ice bath, and sulfuryl chloride (3.58 mL, 44.9mmol) was added dropwise. After a few minutes, the solution was warmedto room temperature for overnight. The mixture was concentrated, dilutedwith ethyl acetate and washed with water. Decolorizing carbon was addedand the mixture was filtered. The resulting solution was dried (sodiumsulfate), filtered and concentrated to afford the title compound (12.4g, 98%). ¹H NMR (CDCl₃) • 7.92 (d, 1H, J=8.5 Hz), 7.57 (d, 1H, J=8.4Hz), 1.57 (s, 9H).

General Procedure for the Synthesis of 7-Sulfoamidebenzoxazoles

2-tert-Butyl-6-chloro-7-(4-methyl-piperazine-1-sulfonyl)-benzooxazole

To a solution of 2-tert-butyl-6-chloro-benzooxazole-7-sulfonyl chloride(2.69 g, 8.73 mmol) and triethylamine (2.44 mL, 17.5 mmol) in THF (60mL) at 0° C. was added 1-methylpiperazine (0.98 mL, 8.83 mmol). Thereaction was warmed to room temperature and allowed to stir overnight.The solution was concentrated and then diluted with water and extactedwith ethyl acetate (3 times). The combined organic layers were driedwith MgSO₄, filtered, and concentrated. Flash chromatography (80% ethylacetate/20% Ethanol) on silica gel gave the title compound (2.45 g,76%). EI-MS m/z 372(M+H)⁺.

General Procedure for the Hydrolysis of the Benzooxazole to the DesiredAniline

6-Amino-3-chloro-2-(4-methyl-piperazine-1-sulfonyl)-phenol

To a solution of2-tert-Butyl-6-chloro-7-(4-methyl-piperazine-1-sulfonyl)-benzooxazole(2.44 g, 6.56 mmol) in 1,4-dioxane (20 mL) was treated with water (4 mL)and conc. H₂SO₄ (4 mL). The mixture was heated to 85° C. for 14 h. Thereaction was cooled to room temperature, and then basified to pH=14 with25% aq NaOH. washed. The mixture was extracted with ethyl acetate (3times), dried with MgSO₄, filtered, and concentrated to afford the titlecompound (1.35 g, 68%). EI-MS m/z 306(M+H)⁺.

The following is the general procedure for thiourea formation:

N-[4-Chloro-2-hydroxy-3[N″,N″-dimethylaminosulfonyl]phenyl]-N′-(2,3-dichlorophenyl)thiourea

A solution of N,N-dimethyl-3-amino-6-chloro-2-hydroxybenzenesulfonamide(356 mg, 1.42 mmol) and 2,3-dichlorophenylisothiocyanate (318 mg, 1.57mmol) in 1.0 mL of N,N-dimethylformamide was stirred at room temperaturefor 3 hours. Purification by column chromatography on silica gel,eluting with ethyl acetate/hexane (30/70, v/v) to give the desiredproduct (440 mg, 68%). EI-MS (m/z) 455.5 (M⁺).

N-[4-Chloro-2-hydroxy-3-[N″,N″-dimethylaminosulfonyl]phenyl]-N′-(2-bromophenyl)thiourea

A solution of N,N-dimethyl-3-amino-6-chloro-2-hydroxybenzenesulfonamide(50 mg, 0.2 mmol) and 2-bromophenylisothiocyanate (43 mg, 0.2 mmol) in0.5 mL of N,N-dimethylformamide was stirred at room temperature for 3hours. Purification by column chromatography on silica gel, eluting withethyl acetate/hexane (30/70, v/v) to give the desired product (66 mg,74%). EI-MS (m/z) 465.5 (M⁺).

The following is the general procedure for protected phenyl thioureaformation:

N-[4-Chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-bromophenyl)thiourea

To a solution ofN-[4-chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-bromophenyl)thiourea(500 mg, 1.07 mmol) in THF (20 mL), tert-butyldimethylsilyl chloride(810 mg, 5.35 mmol) and imidazole (144 mg, 2.14 mmol) were added. Thereaction mixture was stirred at room temperature for 16 hours. Then itwas partitioned between ethyl acetate and water. The combined organicphase was dried and concentrated. Chromatography of the residue onsilica gel (30% Ethyl acetate/Hexane) gave desired product (240 mg, 40%)and recovered starting material (280 mg). EI-MS m/z 580 (M⁺).

N-[4-Chloro2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2,3-dichlorophenyl)thiourea

To a solution ofN-[4-chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2,3dichlorophenyl)thiourea(440 mg, 1.07 mmol) in THF (20 mL), tert-butyldimethylsilyl chloride(730 mg, 4.85 mmol) and imidazole (132 mg, 1.94 mmol) were added. Thereaction mixture was stirred at room temperature for 16 hours. Then itwas partitioned between ethyl acetate and water. The combined organicphase was dried and concentrated. Chromatography of the residue onsilica gel (30% Ethyl acetate/Hexane) gave desired product (270 mg,50%). EI-MS m/z 570 (M⁺).

The following is the general procedure for carbodiimide formation:

N-[4-Chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaninosulfonyl)phenyl]-N′-(2-bromophenyl)carbodiimide

To a solution ofN-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-bromophenyl)thiourea(240 mg, 0.42 mmol) in dichloromethane (10 mL) at 0° C., methanesulfonylchloride (0.065 mL, 0.84 mmol) and triethylamine (0.12 mL, 0.84 mmol)were added. 4-Dimethylaminopyridine was added as a catalyst. Thereaction mixture was stirred at 0° C. for 1 hour, then it waspartitioned between dichloromethane and water. The combined organicphase was dried and concentrated to give the desired product (266.2 mg,crude). ¹H NMR (CDCl₃) δ 0.4 (s, 6H), 1.09 (s, 9H), 2.86 (s, 6H), 7.06(d, 2H), 7.1-7.37 (m, 4H).

N-[4-Chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2,3-dichlorophenyl)carbodiimide

To a solution ofN-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2,3-dichlorophenyl)thiourea(270 mg, 0.45 mmol) in dichloromethane (10 mL) at 0° C., methanesulfonylchloride (0.07 mL, 0.9 mmol) and triethylamine (0.13 mL, 0.9 mmol) wereadded. 4-Dimethylaminopyridine was added as a catalyst. The reactionmixture was stirred at 0° C. for 1 hour, then it was partitioned betweendichloromethane and water. The combined organic phase was dried andconcentrated to give the desired product (270 mg, crude). ¹H NMR (CDCl₃)δ 0.47 (s, 6H), 1.05 (s, 9H), 2.9 (s, 6H), 7.08 (d, 1H), 7.12 (d, 1H),7.2 (t, 1H), 7.31 (m, 2H).

The following is the general procedure for cyanoguanidine formation:

N-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-bromophenyl)cyanoguanidine

To a solution ofN-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-bromophenyl)carbodiimide(266 mg, 0.49 mmol) in acetonitrile (5 mL) at room temperature,cyanamide (83 mg, 1.96 mmol) and N,N-diisopropylethylamine (76 mg, 0.59mmol) was added. The reaction mixture was stirred at room temperaturefor 1 hour. The reaction mixture was concentrated under reducedpressure. The residue was diluted with a mixture of THF (3 mL) andmethanol (1 mL). Cesium fluoride (90 mg, 0.59 mmol) was added at 0° C.The reaction mixture was stirred at 0° C. for 3 hours. The reactionmixture was concentrated under reduced pressure. The residue waspurified by Gilson HPLC to give the desired product (70 mg, 30%). EI-MSm/z 473.75(M⁺).

N-[4-Chloro-2-hydroxy-3-[N″,N″-dimethylaminosulfonyl]phenyl]-N′-(2,3-dichlorophenyl)cyanoguanidine

To a solution ofN-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2,3dichlorophenyl)carbodiimide(270 mg, 0.47 mmol) in acetonitrile (5 mL) at room temperature,cyanamide (79 mg, 1.88 mmol) and N,N-diisopropylethylamine (76 mg, 0.56mmol) was added. The reaction mixture was stirred at room temperaturefor 1 hour. The reaction mixture was concentrated under reducedpressure. The residue was diluted with a mixture of tetrahydrofuran (3mL) and methanol (1 mL). Cesium fluoride (86 mg, 0.56 mmol) was added at0° C. The reaction mixture was stirred at 0° C. for 3 hours. Thereaction mixture was concentrated under reduced pressure. The residuewas purified by Gilson HPLC to give the desired product (98 mg, 48%).EI-MS m/z 473.75(M⁺).

Examples 3, 4 & 5 Preparation ofN-(2-Bromophenyl)-N′-[4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]cyanoguanidine,N-(2,3-Dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]cyanoguanidineandN-(2-Bromophenyl)-N′-[4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]cyanoguanidine

S-(+)-N-(2-Methoxymethyl)pyrrolidin-1-yl-2,6-dichloro-3-nitrobenzenesulfonamide

Following the general procedure for sulfonamide formation outlined inexample 1, 6-dichloro-3-nitrobenzenesulfonyl chloride (1.0 g, 3.44mmol), (S)-(+)-2(methyoxymethyl)pyrrolidine (0.476 mL, 4.13 mmol) andtriethylamine (0.72 mL, 5.16 mmol) were reacted to form the desiredproduct (1.15 g, 91%). EI-MS m/z 368 (M⁻).

S-(+)-N-(2-Methoxymethyl)pyrrolidin-1-yl-6-chloro-2-hydroxy-3-nitrobenzenesulfonamide

Following the general hydrolysis procedure outlined in example 1,S-(+)-N-(2-methoxymethyl)pyrrolidin-1-yl-2,6-dichloro-3-nitrobenzenesulfonamide(1.15 g, 3.12 mmol), 60% sodium hydride (374 mg, 9.36 mmol) and water(73 mg, 4.02 mmol) were reacted to form the desired product (1.0 g,91%). EI-MS m/z 349.1 (M⁻).

S-(+)-N-(2-Methoxymethyl)pyrrolidin-1-yl-3-amino-6-chloro-2-hydroxybenzenesulfonamide

Following the general hydrogenation procedure outlined in example 1,S-(+)-N-(2-methoxymethyl)pyrrolidin-1-yl-6-chloro-2-hydroxy-3-nitrobenzenesulfonamide(1.0 g, 2.86 mmol) was reduced with hydrogen and Pd/C (600 mg) to formthe desired product (0.9 g, 98%). EI-MS m/z 319.1 (M⁻).

N-(2-Bromophenyl)-N′-4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]thiourea

Following the general procedure for thiourea formation outlined inexample 1,S-(+N-(2-methoxymethyl)pyrrolidin-1-yl-3-amino-6-chloro-2-hydroxybenzenesulfonamide(260 mg, 0.85 mmol) and 2-bromophenylisothiocyanate (182 mg, 0.85 mmol)were coupled to form the desired thiourea (231 mg, 53%). EI-MS m/z 535.1(M⁺).

N-(2,3-Dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]thiourea

Following the general procedure for thiourea formation outlined inexample 1,S-(+)-N-(2-methoxymethyl)pyrrolidin-1-yl-3-amino-6chloro-2-hydroxybenzenesulfonamide(407 mg, 1.27 mmol) and 2,3-dichlorophenylisothiocyanate (285 mg, 1.4mmol) were coupled to form the desired thiourea (370 mg, 54%). EI-MS m/z525.1 (M⁺).

N-Phenyl-N′-[4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]thiourea

Following the general procedure for thiourea formation outlined inexample 1,S-(+)-N-(2-methoxymethyl)pyrrolidin-1-yl-3-amino-6-chloro-2-hydroxybenzenesulfonamide(310 mg, 0.97 mmol) and phenylisothiocyanate (144 mg, 1.07 mmol) werecoupled to form the desired thiourea (210 mg, 54%). EI-MS m/z 456 (M⁺).

N-(2-Bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[S-(+)-2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 1,N-(2-bromophenyl)-N′-[4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]thiourea(230 mg, 0.44 mmol), tert-butyldimethylsilyl chloride (332 mg, 2.2 mmol)and imidazole (60 mg, 0.88 mmol) were reacted to form the desiredproduct (136 mg, 48%). EI-MS m/z 650 (M⁺).

N-(2,3-Dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 1,N-(2,3-dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]thiourea(370 mg, 0.71 mmol), tert-butyldimethylsilyl chloride (370 mg, 0.71mmol) and imidazole (97 mg, 1.42 mmol) were reacted to form the desiredproduct (187 mg, 41%). EI-MS m/z 640.2(M⁺).

N-Phenyl-N′-[4-chloro-2-tert-Butyldimethylsilyloxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 1,N-phenyl-N′-4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]thiourea(210 mg, 0.46 mmol), tert-butyldimethylsilyl chloride (349 mg, 2.3 mmol)and imidazole (63 mg, 0.92 mmol) were reacted to form the desiredproduct (125 mg, 41%). EI-MS m/z 571 (M⁺).

N-(2-Bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 1,N-(2-bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]thiourea(136 mg, 0.21 mmol), methanesulfonyl chloride (0.03 mL, 0.42 mmol) andtriethylamine (0.06 mL, 0.42 mmol) were reacted to form the desiredproduct (120 mg, 93%). ¹H NMR (CDCl₃) δ 0.37 (s, 6H), 1.01 (s, 9H), 1.72(m, 4H), 3.25 (s, 3H), 3.3 (m, 2H), 3.45 (m, 2H), 4.15 (m, 1H), 7.10 (d,2H), 7.2 (d, 1H), 7.3 (m, 1H), 7.39 (d, 1H), 7.6 (d, 1H).

N-(2,3-Dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 1,N-(2,3-dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]thiourea(187 mg, 0.3 mmol), methanesulfonyl chloride (0.05 ml, 0.6 mmol) andtriethylamine (0.08 mL, 0.6 mmol) were reacted to form the desiredproduct (160 mg, 90%). ¹H NMR (CDCl₃) δ 0.32 (s, 6H), 1.01 (s, 9H), 1.35(m, 4H), 3.24 (s, 3H), 3.42 (m, 4H), 4.14 (m, 1H), 7.10 (d, 1H), 7.12(d, 1H), 7.2 (t, 1H), 7.29 (d, 1H), 7.32(d, 1H).

N-Phenyl-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 1,N-phenyl-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]thiourea(125 mg, 0.22 mmol), methanesulfonyl chloride (0.04 mL, 0.44 mmol) andtriethylamine (0.06 mL, 0.44 mmol) were reacted to form the desiredproduct (125 mg, crude). ¹H NMR (CDCl₃) δ 0.37 (s, 6H), 1.04 (s, 9H),1.35 (m, 4H), 3.24 (s, 3H), 3.42 (m, 4H), 4.14 (m, 1H), 7.06 (d, 1H),7.16 (d, 1H), 7.19 (m, 2H), 7.25 (d, 1H), 7.35 (t, 2H).

N-(2-Bromophenyl)-N′-[4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 1,N-(2-bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]carbodiimide(152 mg, 0.25 mmol), cyanamide (42 mg, 1.0 mmol) andN,N-diisopropylethylamine (39 mg, 0.3 mmol) were reacted, followed bydesilylation with Cesium fluoride (45.6 mg, 0.3 mmol) to form thedesired product (33 mg, 25%). EI-MS m/z 543.6 (M⁺).

N-(2,3-Dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]cyanoguanidine

Following, the general procedure for carbodiimide formation outlined inexample 1,N-(2,3dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]sulfonylphenyl]carbodiimide(160 mg, 0.26 mmol), cyanamide (44 mg, 1.04 mmol) andN,N-diisopropylethylamine (41 mg, 0.31 mmol) were reacted, followed bydesilylation with Cesium fluoride (48 mg, 0.31 mmol) to form the desiredproduct (35 mg, 25%). EI-MS m/z 532 (M⁺).

N-Phenyl-N′-[4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]cyanoguanidine

Following the general procedure for carbodiimide formation outlined inexample 1,N-phenyl-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]sulfonylphenyl]carbodiimide(125 mg, 0.23 mmol), cyanamide (38.6 mg, 0.92 mmol) andN,N-diisopropylethylamine (36.2 mg, 0.28 mmol) were reacted, followed bydesilylation with Cesium fluoride (42 mg, 0.28 mmol) to form the desiredproduct (38 mg, 35%). LC-MS m/z 462.2.

Examples 6 & 7 Preparation ofN-(2-Bromophenyl)-N′-[4-chloro-2-hydroxy-3-[R-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]cyanoguanidineandN-(2,3-Dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-[R-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]cyanoguanidine

R-N-(2-Methoxymethyl)pyrrolidin-1-yl-2,6-dichloro-3-nitrobenzenesulfonamide

Following the general procedure for sulfonamide formation outlined inexample 1, 6-dichloro-3-nitrobenzenesulfonyl chloride (2.0 g, 6.89mmol), (R)-2-(methyoxymethyl)pyrrolidine (0.783 mL, 8.27 mmol) andtriethylamine (1.44 mL, 10.34 mmol) were reacted to form the desiredproduct (1.69 g, 66%). EI-MS m/z 368 (M⁻).

R-N-(2-Methoxymethyl)pyrrolidin-1-yl-6-chloro-2-hydroxy-3-nitrobenzenesulfonamide

Following the general hydrolysis procedure outlined in example(R)-N-(2-methoxymethyl)pyrrolidin-1-yl-2,6-dichloro-3-nitrobenzenesulfonamide(2.16 g, 5.85 mmol), 60% sodium hydride (702 mg, 17.55 mmol) and water(137 mg, 7.6 mmol) were reacted to form the desired product (2.0 g,97%). EI-MS m/z 349.1 (M⁻).

(R)-N-(2-Methoxymethyl)pyrrolidin-1-yl-3-amino-6-chloro-2-hydroxybenzenesulfonamide

Following the general hydrogenation procedure outlined in example 1,(R)-N-(2-methoxymethyl)pyrrolidin-1-yl-6-chloro-2-hydroxy-3-nitrobenzenesulfonamide(2.0 g, 4.44 mmol) was reduced with hydrogen and Pd/C (1.5 g) to formthe desired product (1.75 g, 96%). EI-MS m/z 319.1 (M⁻).

N-(2-Bromophenyl)-N′-[4-chloro-2-hydroxy-3-[(R)-(2-methoxyl)pyrrolidin-1-yl]aminosulfonylphenyl]thiourea

Following the general procedure for thiourea formation outlined inexample 1,(R)-N-(2-methoxymethyl)pyrrolidin-1-yl-3-amino-6-chloro-2-hydroxybenzenesulfonamide(390 mg, 1.21 mmol) and 2-bromophenylthioisocyanate (286 mg, 1.33 mmol)were coupled to form the desired thiourea (462 mg, 71%). EI-MS m/z 535.1(M⁺).

N-(2,3-Dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-[(R)-2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]thiourea

Following the general procedure for thiourea formation outlined inexample 1,(R)-N-(2-methoxymethyl)pyrrolidin-1-yl-3-amino-6-chloro-2-hydroxybenzenesulfonamide(360 mg, 1.12 mmol) and 2,3-dichlorophenylisothiocyanate (251 mg, 1.23mmol) were coupled to form the desired thiourea (410 mg, 70%). EI-MS m/z525.1 (M⁺).

N-(2-Bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(R)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 1,N-(2-bromophenyl)-N′-[4-chloro-2-hydroxy-3-[(R)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]thiourea(462 mg, 0.86 mmol), tert-butyldimethylsilyl chloride (651 mg, 4.3 mmol)and imidazole (118 mg, 1.73 mmol) were reacted to form the desiredproduct (216 mg, 39%). EI-MS m/z 650 (M⁺).

N-(2,3-Dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(R)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 1,N-(2,3-dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-[(R)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]thiourea(410 mg, 0.78 mmol), tert-butyldimethylsilyl chloride (589 mg, 3.9 mmol)and imidazole (106 mg, 1.56 mmol) were reacted to form the desiredproduct (202 mg, 40%). EI-MS m/z 640.2 (M⁺).

N-(2-Bromophenyl)-N′-[4-chloro2-tert-butyldimethylsilyloxy-3-[(R)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 1,N-(2-bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(R)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]thiourea(216 mg, 0.33 mmol), methanesulfonyl chloride (0.05 mL, 0.66 mmol) andtriethylamine (0.09 mL, 0.66 mmol) were reacted to form the desiredproduct (200 mg, 97%). %). ¹H NMR (CDCl₃) δ 0.37 (s, 6H), 1.02 (s, 9H),1.9 (m, 4H), 3.23 (m, 4H), 3.42 (m, 3H), 4.13 (m, 1H), 7.08 (d, 2H),7.20 (d, 1H), 7.3 (t, 1H), 7.38 (d, 1H), 7.6 (d, 1H).

N-(2,3-Dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(R)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 1,N-(2,3-dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(R)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]thiourea(202 mg, 0.32 mmol), methanesulfonyl chloride (0.05 mL, 0.64 mmol) andtriethylamine (0.09 mL, 0.64 mmol) were reacted to form the desiredproduct (190 mg, 99%). ¹H NMR (CDCl₃) δ 0.34 (s, 6H), 1.04 (s, 9H), 1.4(m, 2H), 1.8 (m, 2H), 3.15 (s, 3H), 3.25 (m, 2H), 3.43 (m, 2H), 4.13 (m,1H), 7.09 (d, 2H), 7.12 (d, 1H), 7.19 (t, 1H), 7.3 (d, 1H), 7.34 (d,1H).

N-(2-Bromophenyl)-N′-[4-chloro-2-hydroxy-3-[(R)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 1,N-(2-bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(R)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]carbodiimide(240 mg, 0.39 mmol), cyanamide (66 mg, 1.56 mmol) andN,N-diisopropylethylamine (63 mg, 0.47 mmol) were reacted, followed bydesilylation with Cesium fluoride (72 mg, 0.47 mmol) to form the desiredproduct (75 mg, 35%). EI-MS m/z 543.6 (M⁺).

N-(2,3-Dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-[(R)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 1,N-(2,3-dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(R)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]carbodiimide(224 mg, 0.37 mmol), cyanamide (63 mg, 1.48 mmol) andN,N-diisopropylethylamine (60 mg, 0.43 mmol) were reacted, followed bydesilylation with Cesium fluoride (67 mg, 0.43 mmol) to form the desiredproduct (70 mg, 36%). EI-MS m/z 533.5 (M⁺).

Examples 8 & 9 Preparation ofN-(2-bromophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-isoxazolidinylaminosulfonylphenyl]cyanoguanidineandN-(2,3-Dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-isoxazolidinylaminosulfonylphenyl]cyanoguanidine

N-(Ethoxycarbonyl)isoxazolidine

To a solution of KOH (6.4 g, 0.11 mol) and hydroxyurethane (12 g, 0.11mol) in ethanol (50 mL) was added 1,3-dibromopropane (5.8 mL, 0.057mol). The resulting suspension was heated at reflux for 1 hour. Afterthe mixture was cooled to room temperature, an additional portion of KOH(3.2 g, 0.055 mol) and of dibromopropane (2.9 mL, 0.028 mol) was added.The mixture was then refluxed for 1 hour, cooled to room temperature,and solvent was evaporated. The residue was suspended in boiling etherthree times and filtered. The combined filtrates were dried over sodiumsulfate, filtered, and evaporated. A portion of 3 g of the crude productwas purified by flush column chromatography (EtOAC/Hexane, gradientelution), yielding 1.18 g of N-(ethoxycarbonyl)isoxazolidine. ¹H NMR(CDCl₃) δ 1.15 (t, 3H), 2.15 (q, 2H), 3.55 (t, 2H), 3.8 (t, 2H), 4.1 (q,2H).

Isoxazolidine Hydrochloride

N-(ethoxycarbonyl)isoxazolidine (1.18 g, 9.1 mmol) was dissolved inaqueous HCl (6N, 7 mL)and heated at reflux for 2 hours. After beingcooled to room temperature, this solution was washed with ether (3×) andthen evaporated affording crude isoxazolidine hydrochloride which wasrecrystalized from ethanol/ether yielding 0.79 g (80%) of isoxazolidinehydrochloride. ¹H NMR (CDCl₃; CH₃OD), δ 2.5 (q, 2H), 3.55 (t, 2H), 4.2(t, 2H).

(N-Isoxazolidinyl)-2.6-dichloro-3-nitrobenzenesulfonamide

Following the general procedure for sulfonamide formation outlined inexample 1, 2,6-dichloro-3-nitrobenzenesulfonyl chloride (1.5 g, 5.2mmol), isoxazolidine hydrochloride (0.56 g, 5.2 mmol) and triethylamine(2.2 mL, 15.5 mmol) were reacted to form the desired product (1.2 g,71%). EI-MS m/z 327 (M⁺).

(N-Isoxazolidinyl)-2-hydroxy-6-dichloro-3-nitrobenzenesulfonamide

Following the general hydrolysis procedure outlined in example 1,(N-isoxazolidinyl)-2,6dichloro-3-nitrobenzenesulfonamide (1.08 g, 3.3mmol), 80% sodium hydride (0.3 g, 10.0 mmol) and water (72 mg, 4.0 mmol)were reacted to form the desired product (0.79 g, 77%). EI-MS m/z 309(M⁺).

(N-Isoxazolidinyl)-2-hydroxy -3-amino-6-dichloro-benzenesulfonamide

Following the general hydrogenation procedure outlined in example 1,(N-isoxazolidinyl)-2-hydroxydichloro-3-nitrobenzenesulfonamide (0.84 g,2.7 mmol) was reduced with hydrogen and Pd/C (840 mg) to form thedesired product (0.75 g crude). EI-MS m/z 279 (M⁺).

N-(2-Bromophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-isoxazolidinylaminosulfonylphenyl]thiourea

Following the general procedure for thiourea formation outlined inexample 1,(N-isoxazolidinyl)-2-hydroxy-3-amino-6-dichloro-benzenesulfonamide (624mg, 2.24 mmol) and 2-bromophenylisothiocyanate (528 mg, 2.46 mmol) werecoupled to form the desired thiourea (620 mg, 56%). EI-MS m/z 493 (M⁺).

N-(2,3-Dichlorophenyl)-N′-[chloro-2-hydroxy-3-(N″-isoxazolidinylaminosulfonylphenyl]thiourea

Following the general procedure for thiourea formation outlined inexample 1,(N-isoxazolidinyl)-2-hydroxy-3-amino-6-dichloro-benzenesulfonamide (590mg, 2.11 mmol) and 2,3-dichlorophenylisothiocyanate (474 mg, 2.53 mmol)were coupled to form the desired thiourea (753 mg, 74%). EI-MS m/z481.75 (M⁻).

N-(2,3-Dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″-isoxazolidinylaminosulfonylphenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 1,N-(2,3-dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-isoxazolidinylaminosulfonylphenyl]thiourea(726 mg, 1.51 mmol), tert-butyldimethylsilyl chloride (1.14 mg, 7.55mmol) and imidazole (205 mg, 3.02 mmol) were reacted to form the desiredproduct (355 mg, 66%). EI-MS m/z 597.83 (M⁺).

N-(2-Bromophenyl)-N′-[chloro-2-tert-butyldimethylsilyloxy-3-(N″-isoxazolidinylaminosulfonylphenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 1,N-(2-bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″-isoxazolidinylaminosulfonylphenyl]thiourea(480 mg, 0.79 mmol), methanesulfonyl chloride (0.12 mL, 1.58 mmol) andtriethylamine (0.22 mL, 1.58 mmol) were reacted to form the desiredproduct (480 mg, crude). ¹H NMR (CDCl₃) δ 0.39 (s, 6H), 1.1 (s, 9H), 2.4(m, 2H), 3.76 (t, 2H), 4.22 (t, 2H), 7.09 (m, 2H), 7.21 (d, 1H), 7.29(m, 1H), 7.45 (d, 1H), 7.6 (d, 1H).

N-(2-,3-Dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″-isoxazolidinylaminosulfonylphenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 1,N-(2,3-dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″-isoxazolidinylaminosulfonylphenyl]thiourea(355 mg, 0.59 mmol), methanesulfonyl chloride (0.09 mL, 1.18 mmol) andtriethylamine (0.16 mL, 1.18 mmol) were reacted to form the desiredproduct (355 mg, crude). ¹H NMR (CDCl₃) δ 0.38 (s, 6H), 1.03 (s, 9H),2.4 (m, 2H), 3.77 (t, 2H), 4.22 (t, 2H), 7.12 (d, 1H), 7.16 (t, 1H),7.26 (d, 1H), 7.31 (d, 1H), 7.40 (d, 1H).

N-(2-Bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″-isoxazolidinylaminosulfonylphenyl]cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 1,N-(2-bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″-isoxazolidinylaminosulfonylphenyl]carbodiimide(480 mg, 0.84 mmol), cyanamide (142 mg, 3.36 mmol) andN,N-diisopropylethylamine (130 mg, 1.0 mmol) were reacted, followed bydesilylation with Cesium fluoride (153.2 mg, 1.0 mmol) to form thedesired product (150 mg, 36%). LC-MS m/z 500.

N-(2-Bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″-isoxazolidinylaminosulfonylphenyl]cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 1,N-(2-,3-dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″-isoxazolidinylaminosulfonylphenyl]carbodiimide(355 mg, 0.63 mmol), cyanamide (105 mg, 2.52 mmol) andN,N-diisopropylethylamine (100 mg, 1.26 mmol) were reacted, followed bydesilylation with Cesium fluoride (115 mg, 0.76 mmol) to form thedesired product (28 mg, 10%). LC-MS m/z 490.

Examples 10 & 11 Preparation ofN-(2-Bromophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-tetrahydroisoxazylaminosulfonyl)phenyl]cyanoguanidineandN-(2,3-Dichlorophenyl)N′-[4-chloro-2-hydroxy-3-(N″-tetrahydroisoxazylaminosulfonyl)phenyl]cyanoguanidine

N-(Ethoxycarbonyl)tetrahydroisoxazine

To a solution of KOH (3.34 g, 59.6 mmol) and hydroxyurethane (6.1 g,58.5 mmol) in ethanol (25 mL) was added 1,4-dibromobutane (3.5mL, 29.3mmol). The resulting suspension was heated at reflux for 1 hour. Afterthe mixture was cooled to room temperature, an additional portion of KOH(1.65 g, 29.4 mmol) and of dibromopropane (1.8 mL, 15 mmol) was added.The mixture was then refluxed for 1 hour, cooled to room temperature,and solvent was evaporated. The residue was suspended in boiling etherthree times and filtered. The combined filtrates were dried over sodiumsulfate, filtered, and evaporated. A portion of 4 g of the crude productwas purified by flash column chromatography (EtOAC/Hexane, gradientelution), yielding 1.85 g of N-(ethoxycarbonyl)tetrahydroisoxazine. ¹HNMR (CDCl₃) δ 1.05 (q, 3H), 1.45 (dd, 2H), 1.55 (dd, 2H), 3.4 (t, 2H),3.7 (t, 2H), 3.95 (q, 2H).

Tetrahydroisoxazine Hydrochloride

N-(ethoxycarbonyl)tetrahydroisoxazine (1.85 g, 11.6 mmol) was dissolvedin aqueous HCl (6N, 7.8 mL)and heated at reflux for 7 hours. After beingcooled to room temperature, this solution was washed with ether (3×) andthen evaporated affording crude tetraisoxazine hydrochloride which wasrecrystalized from ethanol/ether yielding 0.74 g (52%) oftetrahydroisoxazine hydrochloride. ¹H NMR (CH₃OD) δ 1,85 (dd, 2H), 1.95(dd, 2H), 3.4 (t, 2H), 4.25 (t, 2H).

c) (N-Tetrahydroisoxazinyl)-2,6-dichloro-3-nitrobenzenesulfonamide

Following the general procedure for sulfonamide formation outlined inexample 1, 2,6-dichloro-3-nitrobenzenesulfonyl chloride (1.75 g, 6.0mmol) Tetrahydroisoxazine hydrochloride hydrochloride (0.63 g, 5.1 mmol)and triethylamine (2.2 mL, 15.5 mmol) were reacted to form the desiredproduct (1.32 g, 75%). EI-MS m/z 341 (M⁺).

(N-Tetrahydroisoxazinyl)-2-hydroxy-6-chloro-3-nitrobenzenesulfonamide

Following the general hydrolysis procedure outlined in example 1,(N-Tetrahydroisoxazinyl)-2,6-dichloro-3-nitrobenzenesulfonamide (0.1 g,0.29 mmol), 80% sodium hydride (26 mg, 0.88 mmol) and water (6.3 mg,0.35 mmol) were reacted to form the desired product (50 mg, 53%). EI-MSm/z 323 (M⁺).

(N-Tetrahydroisoxazyl)-2-hydroxy-3-amino-6-chlorobenzenesulfonamide

Following the general hydrogenation procedure outlined in example 1,(N-tetrahydroisoxazinyl)-2-hydroxy-6-chloro-3-nitrobenzenesulfonamide(0.76 g, 2.35 mmol) was reduced with hydrogen and Pd/C (760 mg) to formthe desired product (890 mg, crude). EI-MS m/z 293 (M⁺).

N-(2-Bromophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-tetrahydroisoxazylaminosulfonyl)phenyl]thiourea

Following the general procedure for thiourea formation outlined inexample 1,(N-tetrahydroisoxazinyl)-2-hydroxy-3-amino-6-chlorobenzenesulfonamide(620 mg, 2.12 mmol) and 2-bromophenylisothiocyanate (500 mg, 2.33 mmol)were coupled to form the desired thiourea (627 mg, 58%). EI-MS m/z 507(M⁻).

N-(2,3-Dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-tetrahydroisoxazmlaminosulfonyl)phenyl]thiourea

Following the general procedure for thiourea formation outlined inexample 1,(N-tetrahydroisoxazinyl)-2-hydroxy-3-amino-6-chlorobenzenesulfonamide(888 mg, 3.04 mmol) and 2,3-dichlorophenylisothiocyanate (682 mg, 3.34mmol) were coupled to form the desired thiourea (958 mg, 64%). EI-MS m/z495.67 (M⁻).

N-(2-Bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″-tetrahydroisoxazylaminosulfonyl)phenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 1,N-(2-bromophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-tetrahydroisoxazylaminosulfonyl)phenyl]thiourea(627 mg, 1.24 mmol), tert-butyldimethylsilyl chloride (934 mg, 6.2 mmol)and imidazole (169 mg, 2.48 mmol) were reacted to form the desiredproduct (350 mg, 45%). EI-MS m/z 622 (M⁺) .

N-(2,3-Dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″-tetrahydroisoxazylaminosulfonyl)phenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 1,N-(2,3-dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-tetrahydroisoxazylaminosulfonyl)phenyl]thiourea(898 mg, 1.81 mmol), tert-butyldimethylsilyl chloride (1.4 g, 9.05 mmol)and imidazole (246 mg, 3.62 mmol) were reacted to form the desiredproduct (546 mg, 49%). EI-MS m/z 611.81 (M⁺).

N-(2-Bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″-tetrahydroisoxazylaminosulfonyl)phenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 1,N-(2-bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″-tetrahydroisoxazylaminosulfonyl)phenyl]thiourea(385 mg, 0.74 mmol), methanesulfonyl chloride (0.12 mL, 1.48 mmol) andtriethylamine (0.21 mL, 1.48 mmol) were reacted to form the desiredproduct (385 mg, crude). ¹H NMR (CDCl₃) δ 0.35 (s, 6H), 1.02 (s, 9H),1.68 (m, 2H), 1.9 (m, 2H), 3.04 (t, 2H), 4.0 (t, 2H), 7.09 (d, 1H), 7.12(d, 1H), 7.2 (d, 1H), 7.3 (t, 1H), 7.42 (d, 1H), 7.6 (d, 1H).

N-(2,3-Dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″-tetrahydroisoxazylaminosulfonyl)phenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 1,N-(2,3-dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″-tetrahydroisoxazylaminosulfonyl)phenyl]thiourea(547 mg, 0.897 mmol), methanesulfonyl chloride (0.14 mL, 1.79 mmol) andtriethylamine (0.25 mL, 1.79 mmol) were reacted to form the desiredproduct (547 mg, crude). ¹H NMR (CDCl₃) δ 0.36 (s, 6H), 1.03 (s, 9H),1.68 (m, 2H), 1.9 (m, 2H), 3.53 (t, 2H), 4.0 (t, 2H), 7.11 (m, 2H), 7.14(t, 1H), 7.2 (d, 1H), 7.3 (d, 1H), 7.39 (d, 1H).

N-(2-Bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″-tetrahydroisoxazylaminosulfonyl)phenyl]cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 1,N-(2-bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″-tetrahydroisoxazylaminosulfonyl)phenyl]carbodiimide(385 mg, 0.79 mmol), cyanamide (133 mg, 3.16 mmol) andN,N-diisopropylethylamine (127 mg, 0.95 mmol) were reacted, followed bydesilylation with Cesium fluoride (144 mg, 0.95 mmol) to form thedesired product (100 mg, 26%). EI-MS m/z 515 (M⁺).

N-(2,3-Dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″-tetrahydroisoxazylaminosulfonyl)phenyl]cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 1,N-(2,3-dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″-tetrahydroisoxazylaminosulfonyl)phenyl]carbodiimide(547 mg, 0.95 mmol), cyanamide (159 mg, 3.8 mmol) andN,N-diisopropylethylamine (150 mg, 1.14 mmol) were reacted, followed bydesilylation with Cesium fluoride (174 mg, 1.14 mmol) to form thedesired product (190 mg, 40%). LC-MS m/z 504.

Example 12 Preparation ofN-[4-Chloro-2-hydroxy-3-[N″,N″-dimethylaminosulfonylphenyl]-N′-(2-bromophenyl)propylguanidine

N,N-Dimethyl-3-amino-6-chloro-2-hydroxybenzenesulfonamide

To a solution ofN,N-Dimethyl-6-chloro-2-hydroxy-3-nitrobenzenesulfonamide (550 mg, 1.96mmol) in ethyl acetate, was added 10% Pd/C (550 mg). The mixture wasflushed with argon, and then stirred under a hydrogen atmosphere atballoon pressure for 3 hours at room temperature. TLC showed thereaction was not complete. The mixture was filtered through celite andthe celite was washed with methanol. The mixture was retreated under thesame conditions as mentioned above. After 3 hours, the solvent wasevaporated to give the desired product (480 mg, 98%). EI-MS (m/z)250.82, 252.87 (M⁺).

N-[4-Chloro-2-hydroxy-3-[N″,N″-dimethylaminosulfonyl]phenyl]-N′-(2-bromophenyl)thiourea

A solution of N,N-dimethyl-3-amino-6-chloro-2-hydroxybenzene sulfonamide(480 mg, 3.58 mmol) and 2-bromophenylisothiocyanate (0.53 μL, 3.94 mmol)in 10 mL of ethanol was stirred at room temperature for overnight.Purification by column chromatography on silica gel, eluting with ethylacetate/hexane (gradient elution) gave the desired product (368 mg,22%). EI-MS (m/z) 463.67, 465.66, 467.65, 468.64 (M⁺).

N-[4-Chloro-2-tert-Butyldimethylsilyloxy-3-[N″,N″-dimethylaminosulfonyl]phenyl]-N′-(2-bromophenyl)thiourea

To a solution ofN-[4-chloro-2-hydroxy-3-[N″,N″-dimethylaminosulfonyl]phenyl]-N′-2-bromophenyl)thiourea(350 mg, 0.755 mmol) in THF (10 mL), tert-butyldimethylsilyl chloride(171 mg, 1.13 mmol) and iridazole (106 mg, 1.56 mmol) were added. Thereaction mixture was stirred at room temperature for overnight. Then itwas partitioned between ethyl acetate and water. The combined organicphase was dried and concentrated. Chromatography of the residue onsilica gel gave desired product (150 mg, 46%) and recovered startingmaterial (90 mg). EI-MS m/z 577.81, 579.66, 580.65, 581.46, 582.77 (M⁺).

N-[4-Chloro2-tert-butyldimethylsiloxy-3-N″,N″-dimethylaminosulfonyl]phenyl]-N′-(2-bromophenyl)carbodiimide

To a solution ofN-[4-chloro-2-tert-butyldimethylsiloxy-3-[N″,N″-dimethylaminosulfonyl]phenyl]-N′-(2-bromophenyl)thiourea(150 mg, 0.26 mmol) in dichloromethane (5 mL) at 0° C., methanesulfonylchloride (40 μL, 0.52 mmol), 4-Dimethylaminopyridine (10 mg) andtriethylamine (0.11 mL, 0.77 mmol) were added. The reaction mixture wasstirred at 0° C. for 3 hours. Then it was partitioned between ethylacetate and water. The combined organic phase was dried and concentratedto give the desired product (120 mg, 85%). IR: 2140 cm⁻¹.

N-[4-Chloro2-hydroxy-3-[N″,N″-dimethylaminosulfonyl]phenyl]-N′-(2-bromophenyl)propylguanidine

To a solution ofN-[4-chloro-2-tert-butyldimethylsiloxy-3-[N″,N″-dimethylaminosulfonyl]phenyl]-N′-(2-bromophenyl)carbodiimide(60 mg, 0.11 mmol) in tetrahydrofuran (2 mL) at room temperature,N,N-diisopropylethylamine (14 μL, 0.12 mmol) and n-propylamine (10 μL,012 mmol) were added. The reaction mixture was stirred at roomtemperature for 15 minutes, then cooled to 0° C., and TBAF and methanol(1 mL) were added. After 30 minutes, the mixture was quenched withwater, and extracted with ethylacetate. The organic layer wasconcentrated under reduced pressure. The residue was purified by GilsonHPLC to give the desired product (19 mg, 35%). EI-MS m/z 488.7, 490.68,492.67 (M⁺).

Example 13 Preparation ofN-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-chlorophenyl)cyanoguanidine

a)N-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-chlorophenyl)thiourea

Following the general procedure for thiourea formation outlined inexample 12, N,N-dimethyl-3-aminochloro-2-hydroxybenzenesulfonamide (450mg, 1.8 mmol) and 2-chlorophenylisothiocyanate (335.6 mg, 1.98 mmol)were coupled to form the desired thiourea (526 mg, 70%). EI-MS m/z421.47 (M⁺).

b)N-(2-Chlorophenyl)-N[4-[chloro-2-tert-butyldimethylsilyoxy-3-(N″,N″-dimethylaninosulfonyl)phenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 12,N-[4-chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-chlorophenyl)thiourea(500 mg, 1.19 mmol), tert-butyldimethylsilyl chloride (897 mg, 5.95mmol) and imidazole (162 mg, 2.38 mmol) were reacted to form the desiredproduct (311 mg, 49%). EI-MS m/z 535.32 (M⁺).

c)N-(2-Chlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 12,N-(2-chlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]thiourea(311 mg, 0.58 mmol), methanesulfonyl chloride (0.09 mL, 1.16 mmol) andtriethylamine (0.15 mL, 1.16 mmol) were reacted to form the desiredproduct (300 mg, crude). ¹H NMR (CDCl₃) δ 0.37 (s, 6H), 1.03 (s, 9H),2.87 (s, 6H), 7.05 (d, 1H), 7.12 (t, 1H), 7.22 (m, 2H), 7.34 (d, 1H),7.42 (d, 1H).

d)N-(2-Chlorophenyl)-N′-[4-chloro-3-(N″,N″-dimethylaminosulfonyl)phenyl]cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 12,N-(2-chlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]carbodiimide(300 mg, 0.6 mmol), cyanamide (100 mg, 2.4 mmol) andN,N-diisopropylethylamine (94 mg, 0.72 mmol) were reacted, followed bydesilylation with Cesium fluoride (110 mg, 0.72 mmol) to form thedesired product (129 mg, 50%). EI-MS m/z 428.0 (M⁺). ¹H NMR (DMSO-d₆) δ2.86 (s, 6H), 7.2 (d, 1H), 7.32 (t, 1H), 7.36 (t, 1H), 7.43 (d, 1H),7.51 (d, 1H), 7.57 (d, 1H), 8.98 (s, 1H), 9.18 (s, 1H), 10.51 (s, 1H).

Example 14 Preparation ofN-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-fluoro-3-chlorophenyl)cyanoguanidine

a) 2-Fluoro-3-chlorophenylisothiocyanate

Into a solution of 2-fluoro-3-chloroaniline (1.0 g, 6.87 mmol) in 30 mLof toluene at room temperature, thiophosgene (0.8 mL, 10.3 mmol) andtriethylamine (1.12 mL, 8.24 mmol) were added. The mixture was stirredat room temperature for 16 hours. The mixture was parationed betweenethyl acetate and water. The combined organic layer was thenconcentrated to give the desired product (950 mg, 74%). ¹H NMR (CDCl₃) δ7.09 (m, 2H), 7.30 (m, 1H).

b)N-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-fluoro-3-chlorophenyl)thiourea

Following the general procedure for thiourea formation outlined inexample 12, N,N-dimethyl-3-amino-6-chloro-2-hydroxybenzenesulfonamide(500 mg, 2 mmol) and 2-fluoro-3-chlorophenylisothiocyanate (374 mg, 2mmol) were coupled to form the desired thiourea (583 mg, 67%). EI-MS m/z438.2(M⁺).

c)N-(2-Fluoro-3-chlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-fluoro-3-chlorophenyl)thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 12,N-4-chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-fluoro-3-chlorophenyl)thiourea(533 mg, 1.22 mmol), tert-butyldimethylsilyl chloride (913 mg, 6.1 mmol)and imidazole (166 mg, 2.44 mmol) were reacted to form the desiredproduct (412 mg, 61%). EI-MS m/z 552.2 (M⁺).

d)N-(2-Fluoro-3-chlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-fluoro-3-chlorophenyl)carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 12,N-(2-fluoro-3-chlorophenyl)-N′-[4-chloro-2hert-butyldimethylsilyloxy-3-(N″,N″-dmiethylaminosulfonyl)phenyl]-N′-(2-fluoro3-chlorophenyl)thiourea(412 mg, 0.75 mmol), methanesulfonyl chloride (0.14 mL, 1.5 mmol) andtriethylamine (0.24 mL, 1.5 mmol) were reacted to form the desiredproduct (410 mg, crude). ¹H NMR (CDCl₃) δ 0.37 (s, 6H), 0.99 (s, 9H),2.85 (s, 6H), 7.05 (m, 2H), 7.25 (m, 3H).

e) Preparation ofN-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-fluoro-3-chlorophenyl)cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 12,N-[4-chloro-3-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-fluoro3-chlorophenyl)carbodiimide(410 mg, 0.79 mmol), cyanamide (133 mg, 3.16 mmol) andN,N-diisopropylethylamine (122 mg, 0.95 mmol) were reacted, followed bydesilylation with Cesium fluoride (144 mg, 0.95 mmol) to form thedesired product (140 mg, 40%). EI-MS m/z 446.2 (M⁺). ¹H NMR (DMSO-d₆) δ2.87 (s, 6H), 7.2 (d, 1H), 7.32 (t, 1H), 7.40 (m, 2H), 7.59 (d, 1H),9.11 (s, 1H), 9.3 (s, 1H), 10.53 (s, 1H).

Example 15 Preparation ofN-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-trifluoromethylphenyl)cyanoguanidine

a) 2-Trifluoromethylisothiocyanate

Into a solution of 2-trifluoromethylaniline (1.0 g, 6.21 mmol) in 30 mLof toluene at room temperature, thiophosgene (0.72 mL, 9.31 mmol) andtriethylamine (1.01 mL, 7.45 mmol) was added. The mixture was stirred atroom temperature for 16 hours. The mixture was parationed between ethylacetate and water. The combined organic layer was then concentrated togive the desired product (1.01 g, 80%). ¹H NMR (CDCl₃) δ 7.30 (m, 2H),7.54 (t, 1H), 7.64 (d, 1H).

b)N-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-trifluoromethylphenyl)thiourea

Following the general procedure for thiourea formation outlined inexample 12, N,N-dimethyl-3-amino-6-chloro-2-hydroxybenzenesulfonamide(500 mg, 2 mmol) and 2-trifluoromethylphenylisothiocyanate (548 mg, 2mmol) were coupled to form the desired thiourea (469 mg, 52%). EI-MS m/z454.0 (M⁺).

c)N-(2-Trifluoromethylphenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3(N″,N″-dimethylaminosulfonyl)phenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 12,N-[4-chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-trifluoromethylphenyl)thiourea(416 mg, 1.0 mmol), tert-butyldimethylsilyl chloride (750 mg, 5.0 mmol)and imidazole (136 mg, 2.0 mmol) were reacted to form the desiredproduct (250 mg, 45%). EI-MS m/z 568.0 (M⁺).

d)N-(2-Trifluoromethylphenyl)-N′-[chloro-2-tert-Butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 12,N-(2-trifluoromethylphenyl)-N-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaninosulfonyl)phenyl]thiourea(250 mg, 0.44 mmol), methanesulfonyl chloride (0.1 mL, 0.88 mmol) andtriethylamine (0.14 mL, 0.88 mmol) were reacted to form the desiredproduct (250 mg, crude). ¹H NMR (CDCl₃) δ 0.38 (s, 6H), 1.04 (s, 9H),2.87 (s, 6H), 7.07 (d, 1H), 7.2 (d, 1H), 7.29 (m, 2H), 7.54 (t, 1H),7.66 (d, 1H).

e) Preparation ofN-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-trifluoromethylphenyl)cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 12,N-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-trifluoromethylphenyl)carbodiimide(250 mg, 0.47 mmol), cyanamide (79 mg, 1.88 mmol) andN,N-diisopropylethylamine (73 mg, 0.56 mmol) were reacted, followed bydesilylation with Cesium fluoride (86 mg, 0.56 mmol) to form the desiredproduct (80 mg, 37%). EI-MS m/z 462.0 (M⁺). ¹H NMR (DMSO-d₆) δ 2.86 (s,6H), 7.14 (d, 1H), 7.53 (m, 2H), 7.59 (d, 1H), 7.75 (t, 1H), 7.77 (d,1H), 8.91 (s, 1H), 9.28 (s, 1H), 10.53 (s, 1H).

Example 16 Preparation ofN-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-methylphenyl)cyanoguanidine

a) N-[4Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-methylphenyl)thiourea

Following the general procedure for thiourea formation outlined inexample 12, N,N-dimethyl-3-amino-6chloro-2-hydroxybenzenesulfonamide(500 mg, 2 mmol) and 2-methylphenylisothiocyanate (298.4 mg, 2 mmol)were coupled to form the desired thiourea (557 mg, 70%). EI-MS m/z 400.0(M³⁰ ).

b)N-(2-Methylphenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 12,N-[4-Chloro-2-hydroxy-3-[N″,N″-dimethylaminosulfonyl]phenyl]-N′-(2-methylphenyl)thiourea(557 mg, 1.39 mmol), tert-butyldimethylsilyl chloride (1.04 mg, 6.95mmol) and imidazole (189 mg, 2.78 mmol) were reacted to form the desiredproduct (410 mg, 57%). EI-MS m/z 514.2 (M⁺).

c)N-(2-Methylphenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 12,N-(2-methylphenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]thiourea(410 mg, 0.8 mmol), methanesulfonyl chloride (0.14 mL, 1.6 mmol) andtriethylamine (0.23 mL, 1.6 mmol) were reacted to form the desiredproduct (400 mg, crude). ¹H NMR (CDCl₃) δ 0.39 (s, 6H), 1.05 (s, 9H),1.72 (m, 4H), 2.37 (s, 3H), 2.87 (s, 6H), 7.04 (d, 1H), 7.18 (m, 4H),7.29 (d, 1H).

d) Preparation ofN-[4-Chloro2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-methylphenyl)cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 12,N-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-methylphenyl)carbodiimide(400 mg, 0.83 mmol), cyanamide (139.4 mg, 3.32 mmol) andN,N-diisopropylethylamine (129 mg, 1.0 mmol) were reacted, followed bydesilylation with Cesium fluoride (152 mg, 1.0 mmol) to form the desiredproduct (110 mg, 32%). EI-MS m/z 408.2 (M⁺). ¹H NMR (DMSO-d₆) δ 2.22 (s,3H), 2.86 (s, 6H), 7.20 (m, 5H), 7.60 (d, 1H), 8.63(s, 1H), 8.96 (s,1H), 10.50 (s, 1H).

Example 17 Preparation ofN-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-fluorophenyl)cyanoguanidine

a) 2-Fluorophenylisothiocyanate

Into a solution of 2-fluoroaniline (1.0 g, 9.0 mmol) in 30 mL of tolueneat room temperature, thiophosgene (1.06 mL, 13.5 mmol) and triethylamine(1.55 mL, 13.5 mmol) was added. The mixture was stirred at roomtemperature for 16 hours. The mixture was partitioned between ethylacetate and water. The combined organic layer was then concentrated togive the desired product (1.05 g, 76%). ¹H NMR (CDCl₃) δ 7.1-7.3 (m,4H).

b)N-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-fluorolphenyl)thiourea

Following the general procedure for thiourea formation outlined inexample 12, N,N-dimethyl-3-amino-6-chloro-2-hydroxybenzenesulfonamide(500 mg, 2 mmol) and 2-fluorophenylisothiocyanate (306 mg, 2 mmol) werecoupled to form the desired thiourea (497 mg, 62%). EI-MS m/z 404.2(M⁺).

c)N-(2-Fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 12,N-[4-chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-fluorophenyl)thiourea(440 mg, 1.09 mmol), tert-butyldimethylsilyl chloride (817 mg, 5.45mmol) and imidazole (148 mg, 2.18 mmol) were reacted to form the desiredproduct (387 mg, 69%). EI-MS m/z 518.2 (M⁺).

d)N-(2-Fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 12,N-(2-fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]thiourea(357 mg, 0.69 mmol), methanesulfonyl chloride (0.12 mL, 1.38 mmol) andtriethylamine (0.2 mL, 1.38 mmol) were reacted to form the desiredproduct (350 mg, crude). ¹H NMR (CDCl₃) δ 0.38 (s, 6H), 1.05 (s, 9H),1.72 (m, 4H), 2.87 (s, 6H), 7.06 (d, 1H), 7.19 (m, 3H), 7.28 (d, 1H),7.31 (d, 1H).

e) Preparation ofN-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-fluorophenyl)cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 12,N-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-fluorophenyl)carbodiimide(350 mg, 0.72 mmol), cyanamide (121 mg, 2.88 mmol) andN,N-diisopropylethylamine (111 mg, 0.86 mmol) were reacted, followed bydesilylation with Cesium fluoride (132 mg, 0.86 mmol) to form thedesired product (120 mg, 40%). EI-MS m/z 412.2 (M⁺). ¹H NMR (DMSO-d₆) δ2.86 (s, 6H), 7.20 (m, 2H), 7.27 (m, 2H), 7.40 (t, 1H), 7.52 (d, 1H),9.06 (s, 1H), 9.20 (s, 1H), 10.51 (s, 1H).

Example 18 Preparation ofN-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2,3-difluorophenyl)cyanoguanidine

a) 2,3-Difluorophenylisothiocyanate

Into a solution of 2,3-difluoroaniline (1.0 g, 7.74 mmol) in 30 mL oftoluene at room temperature, thiophosgene (0.91 mL, 11.6 mmol) andtriethylamine (1.3 mL, 11.6 mmol) was added. The mixture was stirred atroom temperature for 16 hours. The mixture was partitioned between ethylacetate and water. The combined organic layer was then concentrated togive the desired product (910 mg, 68%). ¹H NMR (CDCl₃) δ 6.98 (m, 2H),7.11 (m, 1H).

b)N-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2,3-difluorophenyl)thiourea

Following the general procedure for thiourea formation outlined inexample 12, N,N-dimethyl-3-amino-6-chloro-2-hydroxybenzenesulfonamide(500 mg, 2 mmol) and 2,3-difluorophenylisothiocyanate (342 mg, 2 mmol)were coupled to form the desired thiourea (467 mg, 54%). EI-MS m/z 422.2(M⁺).

c)N-(2,3-Difluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 12,N-[4-chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2,3-difluorophenyl)thiourea(418 mg, 1.0 mmol), tert-butyldimethylsilyl chloride (745 mg, 5.0 mmol)and imidazole (136 mg, 2.0 mmol) were reacted to form the desiredproduct (347 mg, 65%). EI-MS m/z 536.2 (M⁺).

d)N-(2,3-Difluorophenyl-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 12,N-(2,3-difluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]thiourea(347 mg, 0.69 mmol), methanesulfonyl chloride (0.12 mL, 1.38 mmol) andtriethylamine (0.2 mL, 1.04 mmol) were reacted to form the desiredproduct (340 mg, crude). ¹H NMR (CDCl₃) δ 0.39 (s, 6H), 1.03 (s, 9H),2.86 (s, 6H), 6.95 (t, 1H), 7.03 (m, 2H), 7.08 (d, 1H), 7.31 (d, 1H).

e) Preparation ofN-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2,3-difluorophenyl)cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 12,N-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2,3-difluorophenyl)carbodiimide(340 mg, 0.68 mmol), cyanamide (114 mg, 2.72 mmol) andN,N-disopropylethylamine (106 mg, 0.82 mmol) were reacted, followed bydesilylation with Cesium fluoride (124 mg, 0.82 mmol) to form thedesired product (10 mg, 3.4%). EI-MS m/z 430.0 (M⁺). ¹H NMR (DMSO-d₆) δ2.87 (s, 6H), 7.21 (m, 3H), 7.30 (m, 1H), 7.54 (d, 1H), 9.2 (s, 1H),10.54 (s, 1H).

Example 19 Preparation ofN-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-methyl-3-fluorophenyl)cyanoguanidine

a) 2-Methyl-3-fluorophenylisothiocyanate

Into a solution of 2-methyl-3-fluoroaniline (1.0 g, 8.0 mmol) in 30 mLof toluene at room temperature, thiophosgene (0.94 mL, 12 mmol) andtriethylamine (1.34 mL, 12 mmol) was added. The mixture was stirred atroom temperature for 16 hours. The mixture was partitioned between ethylacetate and water. The combined organic layer was then concentrated togive the desired product (1.1 g, 82%). EI-MS m/z 168.2 (M⁺).

b)N-[4Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-2-methyl-3-fluorophenyl)thiourea

Following the general procedure for thiourea formation outlined inexample 12, N,N-dimethyl-3-amino-6-chloro-2-hydroxybenzenesulfonamide(560 mg, 2.23 mmol) and 2-methyl-3-fluorophenylisothiocyanate (372 mg,2.23 mmol) were coupled to form the desired thiourea (570 mg, 61%).EI-MS m/z 418.2 (M⁺).

c)N-(2-Methyl-3-fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 12,N-[4-chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-methyl-3-fluorophenyl)thiourea(530 mg, 1.27 mmol), tert-butyldimethylsilyl chloride (951 mg, 6.35mmol) and imidazole (173 mg, 2.54 mmol) were reacted to form the desiredproduct (331 mg, 49%). EI-MS m/z 532.2 (M⁺).

d)N-(2-Methyl-3-fluorophenyl-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 12,N-(2-methyl-3-fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]thiourea(330 mg, 0.62 mmol), methanesulfonyl chloride (0.12 mL, 1.38 mmol) andtriethylamine (0.2 mL, 1.04 mmol) were reacted to form the desiredproduct (320 mg, crude). ¹H NMR (CDCl₃) δ 0.38 (s, 6H), 1.05 (s, 9H),2.28 (s, 3H), 2.85 (s, 6H), 6.89 (t, 1H), 6.93 (d, 1H), 7.05 (d, 1H),7.13 (m, 1H), 7.19 (d, 1H).

f) Preparation ofN-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-methyl-3-fluorophenyl)cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 12,N-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-methyl-3-fluorophenyl)carbodiimide(320 mg, 0.64 mmol), cyanamide (108 mg, 2.56 mmol) andN,N-diisopropylethylamine (99 mg, 0.77 mmol) were reacted, followed bydesilylation with Cesium fluoride (117 mg, 0.77 mmol) to form thedesired product (120 mg, 44%). EI-MS m/z 426.2 (M⁺). ¹H NMR (DMSO-d₆) δ2.12 (s, 3H), 2.86 (s, 6H), 7.09 (m, 3H), 7.22 (m, 1H), 7.56 (d, 1H),8.82 (s, 1H), 9.10 (s, 1H), 10.51 (s, 1H).

Example 20 Preparation ofN-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-chloro-3-fluorophenyl)cyanoguanidine

a) 2-Chloro-3-fluorobenzoic Acid

A solution of 3-fluorobenzoic acid (8.0 g, 64.3 mmol) in 40 mL of THFwas added dropwise to a solution of sec-butyllithium (90 mL, 128.6 mmol)and N,N,N′,N′-tetramethylethylenediamine (20.0 mL, 147.9 mmol) in THF(100 mL) at −90° C. After addition, the reaction mixture was stirred at−90° C. for 30 minutes. Hexachloroethane (54 g, 257.2 mmol) in THF (100mL) was added to reaction mixture dropwise. Then the reaction mixturewas stirred at −78° C. to room temperature for 16 hours. The solvent wasevaporated and the water was added to the residue. The reaction mixturewas acidifed to PH=1 by added conc. hydrochloric acid. Then the reactionmixture was extracted with ether (3×). The combined organic phase wasdried and conc. The crude product was washed with hexane for 3 times.Then it was filtered to get pure product (9.2 g, 93%). EI-MS m/z 172.89(M⁻).

b) 2-Chloro-3-fluoroaniline

To a solution of 2-chloro-3-fluorobenzoic acid (4 g, 23 mmol) in CHCl₃(50 mL), sulfonic acid (64 mL) was added and then sodium azide (2.62 g,1.75 mmol) was added by portion. After addition, the reaction mixturewas stirred at 50° C. for 16 hours. The solvent was evaporated andbasicified with ammonium hydroxide at ice-bath. Then was extracted withethyl acetate (3×). The combine organic phase was dried and conc. togive the desired product (2.61 g, 78%). LC-MS m/z 146.2 (M⁺).

c) 2-Chloro-3-fluorophenylisothiocyanate

Into a solution of 2-chloro-3-fluoroaniline (2.61 g, 17.94 mmol) in 50mL of toluene at room temperature, thiophosgene (2.1 mL, 26.91 mmol) andtriethylamine (3.02 mL, 26.91 mmol) was added. The mixture was stirredat room temperature for 16 hours. The mixture was partitioned betweenethyl acetate and water. The combined organic layer was thenconcentrated to give the desired product (2.99 g, 89%). ¹H NMR (CDCl₃) δ7.10 (m, 1H), 7.22 (m, 1H).

d)N-[4Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-chloro-3-fluorophenyl)thiourea

Following the general procedure for thiourea formation outlined inexample 12, N,N-dimethyl-3-amino-6-chloro-2-hydroxybenzenesulfonamide(500 mg, 2.0 mmol) and 2chloro-3-fluorophenylisothiocyanate (374 mg, 2.0mmol) were coupled to form the desired thiourea (623 mg, 71%). LC-MS m/z438.2 (M⁺).

e)N-(2-Chloro-3-fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 12,N-[4-chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-chloro-3-fluorophenyl)thiourea(575 mg, 1.32 mmol), tert-butyldimethylsilyl chloride (991 mg, 6.6 mmol)and imidazole (179 mg, 2.64 mmol) were reacted to form the desiredproduct (367 mg, 50%). LC-MS m/z 552.2 (M⁺).

f)N-(2-Chloro-3-fluorophenyl-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 12,N-(2-chloro-3-fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]thiourea(367 mg, 0.67 mmol), methanesulfonyl chloride (0.12 mL, 1.38 mmol) andtriethylamine (0.2 mL, 1.04 mmol) were reacted to form the desiredproduct (360 mg, crude). ¹H NMR (CDCl₃) δ 0.36 (s, 6H), 1.03 (s, 9H),1.2.87 (s, 6H), 7.01 (m, 2H), 7.09 (d, 1H), 7.21 (m, 1H), 7.34 (d, 1H).

g) Preparation ofN-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-chloro-3-fluorophenyl)cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 12,N-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-chloro-3-fluorophenyl)carbodiimide(360 mg, 0.7 mmol), cyanamide (118 mg, 2.8 mmol) andN,N-diisopropylethylamine (109 mg, 0.84 mmol) were reacted, followed bydesilylation with Cesium fluoride (128 mg, 0.84 mmol) to form thedesired product (120 mg, 39%). LC-MS m/z 446.2 (M⁺). ¹H NMR (DMSO-d₆) δ2.86 (s, 6H), 7.20 (d, 1H), 7.33 (t, 1H), 7.40 (m,2H), 7.58 (d, 1H),9.11 (s, 1H), 9.31 (s, 1H), 10.52 (s, 1H).

Example 21 Preparation ofN-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-chloro4-fluorophenyl)cyanoguanidine

a) 2-Chloro-4-fluorophenylisothiocyanate

Into a solution of 2-chloro-4fluoroaniline (500 mg, 3.44 mmol) in amixture of chloroform and water (10 mL/10 mL) at room temperature,thiophosgene (0.53 mL, 6.88 mmol) and sodium bicarbonate (1.09 g, 10.32mmol) were added. The mixture was stirred at room temperature for 16hours. The mixture was partitioned between chloroform and water. Thecombined organic layer was then concentrated to give the desired product(586 mg, 91%). ¹H NMR (CDCl₃) δ 6.98 (m, 1H), 7.20 (m, 2H).

b)N-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2chloro-4-fluorophenyl)thiourea

Following the general procedure for thiourea formation outlined inexample 12, N,N-dimethyl-3-amino-6-chloro-2-hydroxybenzenesulfonamide(526 mg, 2.10 mmol) and 2-chloro-4-fluorophenylisothiocyanate (586 mg,3.1 mmol) were coupled to form the desired thiourea (505 mg, 55%). EI-MSm/z 437.75 (M⁻).

c)N-(2-Chloro-4-fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 12,N-[4-chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-chlorofluorophenyl)thiourea(470 mg, 1.08 mmol), tert-butyldimethylsilyl chloride (810 mg, 5.4 mmol)and imidazole (147 mg, 2.16 mmol) were reacted to form the desiredproduct (420 mg, 71%). EI-MS m/z 551.75 (M⁺).

d)N-(2-Chloro-4-fluorophenyl-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 12,N-(2-chloro4-fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]thiourea(420 mg, 0.76 mmol), methanesulfonyl chloride (0.11 mL, 1.52 mmol) andtriethylamine (0.22 mL, 1.52 mmol) were reacted to form the desiredproduct (420 mg, crude). ¹H NMR (CDCl₃) δ 0.36 (s, 6H), 1.02 (s, 9H),2.89 (s, 6H), 6.99 (m, 1H), 7.08 (d, 1H), 7.19 (m, 2H), 7.33 (d, 1H).

e) Preparation ofN-[4-Chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2-chloro-4-fluorophenyl)cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 12,N-[4-chloro-2-tert-butyldimethylsilyloxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]N′-(2-chloro4-fluorophenyl)carbodiimide(420 mg, 0.81 mmol), cyanamide (136 mg, 3.24 mmol) andN,N-diisopropylethylamine (126 mg, 1.62 mmol) were reacted, followed bydesilylation with Cesium fluoride (148 mg, 1.62 mmol) to form thedesired product (180 mg, 50%). EI-MS m/z 446.2 (M⁺).

¹H NMR (CDCl₃) δ 2.86 (s, 6H), 7.16 (d, 1H), 7.24 (t, 1H), 7.46 (m, 1H),7.55 (m, 2H), 8.94 (s, 1H), 9.15 (s, 1H), 10.50 (s, 1H).

Example 22 Preparation ofN-(2-Chloro-4-fluorophenyl)-N′-[4-chloro-2hydroxy-3-(pipridone-4-ketone)aminosulfonylphenyl]cyanoguanidine

a)N-(2-Chloro-4-fluorophenyl)-N′-[4-chloro-2-hydroxy-3-(piperidone-4-ketone)aminosulfonylphenyl]thiourea

Following the general procedure for thiourea formation outlined inexample 12,(N-piperidone-4-ketone)-2-hydroxy-3-amino-6-dichloro-benzenesulfonamide(732 mg, 2.4 mmol) and 2-chloro-4-fluorophenylisothiocyanate (seeExample 46, 500 mg, 2.67 mmol) were coupled to form the desired thiourea(704 mg, 54%). EI-MS m/z 491.96 (M⁻).

b)N-(2-Chloro-4-fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(piperidone-4-ketone)aminosulfonylphenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 12,N-(2-chlorofluorophenyl)-N′-[4-chloro-2-hydroxy-3-(piperidone-4-ketone)aminosulfonylphenyl]thiourea(704 mg, 1.44 mmol), tert-butyldimethylsilyl chloride (1.08 g, 7.2 mmol)and imidazole (196 mg, 2.88 mmol) were reacted to form the desiredproduct (340 mg, 39%). LC-MS m/z 606.2 (M⁺).

c)N-(2-Chloro-4-fluorophenyl)-N′-[chloro-2-tert-butyldimethylsilyloxy-3-(piperidone-4-ketone)aminosulfonylphenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 12,N-(2-chloro-4-fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(piperidone4-ketone)aminosulfonylphenyl]thiourea(340 mg, 0.56 mmol), methanesulfonyl chloride (0.08 mL, 1.12 mmol) andtriethylamine (0.16 mL, 1.12 mmol) were reacted to form the desiredproduct (330 mg, crude). LC-MS m/z 572.2 (M⁺).

d)N-(2-Chloro-4-fluorophenyl)-N′-[4-chloro-2-hydroxy-3-(piperidone-4-ketone)aminosulfonylphenyl]cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 12,N-(2-chloro4fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(piperidone-4-ketone)aminosulfonylphenyl]carbodiimide(330 mg, 0.58 mmol), cyanamide (104 mg, 2.48 mmol) andN,N-diisopropylethylamine (96 mg, 0.74 mmol) were reacted, followed bydesilylation with Cesium fluoride (113 mg, 0.74 mmol) to form thedesired product (76 mg, 26%). LC-MS m/z 500.2. ¹H NMR (DMSO-d₆) δ 2.44(t, 4H), 3.64 (s, 4H), 7.19 (d, 1H), 7.25 (t, 1H), 7.46 (m, 1H), 7.58(d, 2H), 8.93 (s, 1H), 9.20 (s, 1H), 10.53 (s, 1H).

Example 23 Preparation ofN-(2,3-Dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-(piperidone-4-ketone)aminosulfonylphenyl]cyanoguanidine

a)N-(2,3-Dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-(piperidone-4-ketone)aminosulfonylphenyl]thiourea

Following the general procedure for thiourea formation outlined inexample 12,(N-piperidone4-ketone)-2-hydroxy-3-amino-6-dichloro-benzenesulfonamide(1.0 g, 3.3 mmol) and 2,3-dichlorophenylisothiocyanate (669 mg, 3.3mmol) were coupled to form the desired thiourea (500 mg, 30%). LC-MS m/z508.2.

b)N-(2,3-Dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(piperidone-4-ketone)aminosulfonylphenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 12,N-(2,3-dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-(piperidone-4-ketone)aminosulfonylphenyl]thiourea(500 mg, 0.98 mmol), tert-butyldimethylsilyl chloride (765 mg, 4.9 mmol)and imidazole (140 mg, 1.98 mmol) were reacted to form the desiredproduct (289 mg, 39%). LC-MS m/z 622.2 (M⁺).

c)N-(2,3-Dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(piperidone-4-ketone)aminosulfonylphenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 12,N-(2,3-dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(piperidone-4-ketone)aminosulfonylphenyl]thiourea(289 mg, 0.46 mmol), methanesulfonyl chloride (0.07 mL, 0.92 mmol) andtriethylamine (0.13 mL, 0.92 mmol) were reacted to form the desiredproduct (280 mg, crude). ¹H NMR (CDCl₃) δ 0.36 (s, 6H), 1.03 (s, 9H),2.54 (t, 4H), 3.63 (t, 4H), 7.09 (d, 1H), 7.1-7.27 (m, 2H), 7.32 (d,1H), 7.38 (d, 1H).

d)N-(2,3-Dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-(piperidone-4-ketone)aminosulfonylphenyl]cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 12,N-(2,3-dichlorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(piperidone-4-ketone)aminosulfonylphenyl]carbodiimide(280 mg, 0.48 mmol), cyanamide (80.6 mg, 1.92 mmol) andN,N-diisopropylethylamine (75 mg, 0.57 mmol) were reacted, followed bydesilylation with Cesium fluoride (88 mg, 0.57 mmol) to form the desiredproduct (40 mg, 16%). LC-MS m/z 500.2. ¹H NMR (DMSO-d₆) δ 2.44 (t, 4H),3.64 (s, 4H), 7.13 (m, 1H), 7.41 (m, 2H), 7.54 (m, 2H), 7.68 (d, 1H),9.07 (s, 1H), 9.4 (s, 1H), 10.56 (s, 1H).

Example 24 Preparation ofN-(2-Chloro-3-fluorophenyl)-N′-[4-chloro-2-hydroxy-3-(piperidone-4ketone)aminosulfonylphenyl]cyanoguanidine

a)N-(2-Chloro-3-fluorophenyl)-N′-[4-chloro-2-hydroxy-3-(piperidone-4-ketone)aminosulfonylphenyl]thiourea

Following the general procedure for thiourea formation outlined inexample12,(N-piperidone-4-ketone)-2-hydroxy-3-amino-6-dichloro-benzenesulfonamide(908 mg, 3.3 mmol, not clean) and 2-chloro-3-fluorophenyllisothiocyanate(See Example 45, 620 mg, 3.3 mmol) were coupled to form the desiredthiourea (350 mg, 24%). LC-MS m/z 492.2.

b)N-(2-Chloro-3-fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(piperidone-4-ketone)aminosulfonylphenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 12,N-(2-chloro-3-fluorophenyl)-N′-[4-chloro-2-hydroxy-3-(piperidone-4-ketone)aminosulfonylphenyl]thiourea(350 mg, 0.71 mmol), tert-butyldimethylsilyl chloride (535 mg, 3.55mmol) and imidazole (98 mg, 1.42 mmol) were reacted to form the desiredproduct (280 mg, 65%). LC-MS m/z 606.2 (M⁺).

c)N-(2-Chloro-3-fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(piperidone-4-ketone)aminosulfonylphenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 12,N-(2-chloro-3-fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(piperidone-4-ketone)aminosulfonylphenyl]thiourea(280 mg, 0.46 mmol), methanesulfonyl chloride (0.07 mL, 0.92 mmol) andtriethylamine (0.13 mL, 0.92 mmol) were reacted to form the desiredproduct (280 mg, crude). ¹H NMR (CDCl₃) δ 0.36 (s, 6H), 1.03 (s, 9H),2.54 (t, 4H), 3.63 (t, 4H), 7.01 (d, 1H), 7.11 (d, 1H), 7.24 (m, 2H),7.36 (d, 1H).

d)N-(2-Chloro-3-fluorophenyl)-N′-[4-chloro-2-hydroxy-3-(piperidone-4-ketone)aminosulfonylphenyl]cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 12,N-(2-chloro-3-fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(piperidone-4-ketone)aminosulfonylphenyl]carbodiimide(280 mg, 0.49 mmol), cyanamide (82.3 mg, 1.96 mmol) andN,N-diisopropylethylamine (75 mg, 0.57 mmol) were reacted, followed bydesilylation with Cesium fluoride (88 mg, 0.57 mmol) to form the desiredproduct (42 mg, 16%). LC-MS m/z 500.2. ¹H NMR (DMSO-d₆) δ 2.44 (t, 4H),3.64 (s, 4H), 7.2 (m, 1H), 7.22 (t, 1H), 7.34 (m, 3H), 7.58 (d, 1H), 9.1(s, 1H), 9.3 (s, 1H), 10.57 (s, 1H).

Example 25 Preparation ofN-(2-Bromo-3-fluorophenyl)-N′-[4-chloro-2-hydroxy-3-(piperidone-4-ketone)aminosulfonylphenyl]cyanoguanidine

a)N-(2-Broom-3-fluorophenyl)-N′-[4-chloro-2-hydroxy-3-(piperidone-4-ketone)aminosulfonylphenyl]thiourea

Following the general procedure for thiourea formation outlined inexample12,(N-piperidone-4-ketone)-2-hydroxy-3-amino-6-dichloro-benzenesulfonamide(1.5 g, 4.92 mmol, not clean) and 2-bromo-3-fluorophenyllisothiocyaniate(See Example 16, 1.0 g, 4.92 mmol) were coupled to form the desiredthiourea (550 mg, 24%). EL-MS m/z 535.96 (M⁺).

b)N-(2-Bromo-3-fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(piperidone-4-ketone)aminosulfonylphenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 12,N-(2-bromo-3-fluorophenyl)-N′-[4-chloro-2-hydroxy-3-(piperidone-4-ketone)aminosulfonylphenyl]thiourea(550 mg, 1.03 mmol), tert-butyldimethylsilyl chloride (773 mg, 5.15mmol) and imidazole (142 mg, 2.06 mmol) were reacted to form the desiredproduct (256 mg, 38%). LC-MS m/z 649.96 (M⁻).

c)N-(2-Bromo-3-fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(piperidone4-ketone)aminosulfonylphenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 12,N-(2-bromo-3-fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-(piperidone-4-ketone)aminosulfonylphenyl]thiourea(256 mg, 0.4 mmol), methanesulfonyl chloride (0.07 mL, 0.92 mmol) andtriethylamine (0.13 mL, 0.92 mmol) were reacted to form the desiredproduct (256 mg, crude).). ¹H NMR (CDCl₃) δ 0.36 (s, 6H), 1.03 (s, 9H),2.54 (t, 4H), 3.63 (t, 4H), 6.96 (t, 1H), 7.0 (d, 1H), 7.10 (d, 1H),7.24 (m, 1H), 7.39 (d, 1H).

d)N-(2-Bromo-3-fluorophenyl)-N′-[4-chloro-2-hydroxy-3-(piperidone-4-ketone)aminosulfonylphenyl]cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 12,N-(2-bromo-3-fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsiyloxy-3-(piperidone-4-ketone)aminosulfonylphenyl]carbodiimide(256 mg, 0.42 mmol), cyanamide (82.3 mg, 1.96 mmol) andN,N-disopropylethylamine (75 mg, 0.57 mmol) were reacted, followed bydesilylation with Cesium fluoride (88 mg, 0.57 mmol) to form the desiredproduct (48 mg, 21%). LC-MS m/z 544.2. ¹H NMR (DMSO-d₆) δ 2.44 (t, 4H),3.64 (s, 4H), 7.10 (m, 1H), 7.28 (m, 2H), 7.42 (m, 1H), 7.59 (m, 1H),9.02 (s, 1H), 9.53 (s, 1H), 10.5 (s, 1H).

Example 26 Preparation ofN-(2-Bromophenyl)-N′-4-chloro-2-hydroxy-3-(homopiperazineaminosulfonylphenyl)phenyl]cyanoguanidine

a) N-(3,4Dichlorophenyl)-2,2-dimethyl-propionamide

3,4-dichloroaniline (150 g) in TBME (1 L) was cooled to 10-15° C. 30% aqNaOH (141 g, 1.14 equiv) was added, and the solution stirred vigorouslyvia overhead mechanical stirrer. Trimethylacetyl chloride (“PivCl”, 126mL) was added at such a rate as to keep the internal temperature below30° C. During this addition, the solution mixture becomes thick withwhite solid product. When the addition was complete (10-15 min), themixture was heated to 30-35° C. for 1 hr, and then allowed to cool. Thereaction mixture was held at −5° C. (overnight), and then filtered,rinsing first with 90:10 water/MeOH (600 mL) and then water (900 mL).Drying under vacuum yielded 195 g (86%) product, as off-white crystals.LCMS m/z 246(M−H)⁺.

b) 2-tert-Butyl-6-chloro-benzooxazole-7-sulfonyl Chloride

The solution of N3,4dichloro-phenyl)-2,2dimethyl-propionamide (10 g, 41mmol) in dry THF (100 mL) was cooled to −72° C. under argon. n-Butyllithium (1.6M in hexane, 64 mL, 102 mmol) was added dropwise. Thesolution warmed to ca. −50° C. over 45 minutes, and then was kept in the−25-−10° C. range for 2 hrs. The solution was then recooled to −78° C.,and sulfur dioxide was bubbled through the solution for 30 min. Thesolution was then allowed to warm to room temperature for 2 h, and a Arstream was bubbled through the solution, with a gas outlet provided sothat any excess sulfur dioxide could escape during the warming. The THFsolution was cooled in an ice bath, and sulfuryl chloride (3.58 mL, 44.9mmol) was added dropwise. After a few minutes, the solution was warmedto room temperature for overnight. The mixture was concentrated, dilutedwith ethyl acetate and washed with water. Decolorizing carbon was addedand the mixture was filtered. The resulting solution was dried (sodiumsulfate), filtered and concentrated to afford the title compound (12.4g, 98%). ¹H NMR (CDCl₃) • 7.92 (d, 1H, J=8.5 Hz), 7.57 (d, 1H, J=8.4Hz), 1.57 (s, 9H).

c) Homopiperazine-carboxylic Acid tert-Butyl Ester

To a solution of homopiperazine (5.0 g, 49.92 mmol) in dichloromethane(100 mL), Di-tert-butyl dicarbonate (3.63 g, 16.64 mmol) andtriethylamine (6.96 mL, 49.92 mmol) were added at room temperature. Thenthe reaction mixture was stirred at room temperature for 16 hours. Thesolid was filtered and the organic phase was washed with water (3×). Theorganic phase was dried (sodium sulfate), filtered and concentrated toafford the title compound (6.5 g, 66%). EI-MS m/z 197.79(M⁻).

General Procedure for the Synthesis of Sulfonylamides

d) 2-tert-Butyl-6-chloro-7-(homopiperazine-1-sulfonyl)-benzooxazole

To a solution of 2-tert-butyl-6-chloro-benzooxazole-7-sulfonyl chloride(5.54, 18 mmol) and triethylamine (2.51 mL, 18 mmol) in THF (100 mL) at0° C. was added homopiperazine-carboxylic acid tert-butyl ester (3.0 g,15 mmol). The reaction was warmed to room temperature and allowed tostir overnight. The solution was concentrated and then diluted withwater and extacted with ethyl acetate (3 times). The combined organiclayers were dried with MgSO₄, filtered, and concentrated. Flashchromatography (80% ethyl acetate/20% Ethanol) on silica gel gave thetitle compound (4.0 g, 61%). LC-MS m/z 473.2.

General Procedure for the Hydrolysis of the Benzooxazole to the DesiredAniline

e) 6-Amino-3-chloro-2-(homopiperazine-1-sulfonyl)-phenol

To a solution of2-tert-Butyl-6-chloro-7-(homopiperazine-1-sulfonyl)-benzooxazole (2.0 g,4.24 mmol) in 1,4-dioxane (56 mL) was treated with water (3.5 mL) andconc. H₂SO₄ (3.5 mL). The mixture was heated to 100° C. for 16 h. Thereaction was cooled to room temperature, and then basified to pH=14 with25% aq NaOH. washed. The mixture was extracted with ethyl acetate (3times), dried with MgSO₄, filtered, and concentrated to afford the titlecompound (1.04 g, 80%). EI-MS m/z 306.2(M⁺).

General Procedure for the Protected Amine Group

f)6-Amino-3-chloro-2-[(4-N-9-fluorenylmethylformate)homopiperazine-1-sulfonyl)]-phenol

To a solution of 6-Amino-3-chloro-2-(homopiperazine-1-sulfonyl)-phenol(1.0 g, 3.27 mmol) in 1,4-dioxane (30 mL), 10% of sodium carbonate (7.5mL) was added. Then the reaction mixture was cooled down to 0° C.,9-fluorenylmethyl chloroformate (800 mg, 3.27 mmol) was added. Afteraddition, the reaction mixture was stirred at 0° C. for 1 hour. Thesolvent was evaporated. The residue was partitioned between ethylacetate and 10% of sodium carbonate. The combined organic phase wasdried with MgSO₄, filtered, and concentrated to afford the tide compound(2.0 g, crude). EI-MS m/z 528.2(M⁺).

g)N-(2-Bromophenyl)-N′-[4-chloro-2-hydroxy-3-[(4N-9-fluorenylmethylformate)homopiperazineaminosulfonylphenyl]thiourea

Following the general procedure for thiourea formation outlined inexample 12,6-Amino3-chloro-2-[(4N-9-fluorenylmethylformate)homopiperazine-1-sulfonyl)]-phenol(2.0 g, 3.79 mmol, not clean) and 2-bromophenylisothiocyanate (811 mg,3.79 mmol) were coupled to form the desired thiourea (2.0 g, 71%). EI-MSm/z 740.61 (M⁻).

h)N-(2-Bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(4-N-9-fluorenylmethylformate)homopiperazineAminosulfonylphenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 12,N-(2-bromophenyl)-N′-[4-chloro-2-hydroxy-3-[(4-N-9-fluorenylmethylformate)homopiperazineaminosulfonylphenyl]thiourea (1.0 g, 1.35 mmol), tert-butyldimethylsilylchloride (1.02 g, 6.75 mmol) and imidazole (184 mg, 2.7 mmol) werereacted to form the desired product (836 mg, 73%). EI-MS m/z 854.97(M⁻).

i)N-(2-Bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(4-N-9-fluorenylmethylformate)homopiperazineAminosulfonylphenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 12,N-(2-bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(4-N-9-fluorenylmethylformate)homopiperazineaminosulfonylphenyl]thiourea (836 mg, 0.98 mmol), methanesulfonylchloride (0.15 mL, 1.96 mmol) and triethylamine (0.27 mL, 1.96 mmol)were reacted to form the desired product (772 mg, crude). ¹H NMR (CDCl₃)δ 0.40 (s, 6H), 1.06 (s, 9H), 1.58 (t, 1H), 1.68 (m, 1H), 1.92 (t, 4H),3.03 (q, 1H), 3.10 (t, 1H), 3.36 (m, 3H), 3.44 (m, 2H), 4.23 (t, 1H),4.64 (d, 2H), 7.07 (d, 1H), 7.33 (m, 8H), 7.60 (m, 3H), 7.73 (m, 2H).

j) N-(2-Bromophenyl)-N′-[4-chloro-2-hydroxy-3-(homopiperazineAminosulfonyl)phenyl]cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 12,N2-bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(4-N-9-fluorenylmethylformate)homopiperazineaminosulfonylphenyl]carbodiimide (256 mg, 0.42 mmol), cyanamide (82.3mg, 1.96 mmol) and N,N-diisopropylethylamine (75 mg, 0.57 mmol) werereacted, followed by desilylation with Cesium fluoride (88 mg, 0.57mmol) and deprotected amino with 20% piperidine (8 mL) in THF (40 mL) atroom temperature for 30 minutes to form the desired product. The desiredproduct (250 mg) was purified by Gilson HPLC to give pure product (50mg, 20%). LC-MS m/z 527.2. ¹H NMR (DMSO-d₆) δ 1.68 (m, 2H), 3.06 (m,2H), 3.2 (m, 2H), 3.34 (m, 2H), 3.71 (m, 2H), 6.25 (m, 1H), 7.19 (m,1H), 7.4 (m, 3H), 7.71 (m, 1H), 8.76 (s, 1H).

Example 27 Preparation of N-(2-Bromophenyl)-N′-[4-chloro-2-hydroxy-3-(N-methylhomopiperazine-aminosulfonylphenyl)phenyl]cyanoguanidine

To a solution ofN-(2-bromophenyl)-N′-[4-chloro-2-hydroxy-3-(homopiperazine-aminosulfonylphenyl)phenyl]cyanoguanidine(287 mg, 0.54 mmol) in diglyme (10 mL), paraformalde (35 mg, 1.08 mmol)and titanium isoproxide (0.17 mL, 0.54 mmol) were added at roomtemperature. The reaction mixture was stirred at 60° C. for 1 hour andstirred at room temperature for 30 minutes. Then sodium borohydride (22mg, 0.65 mmol) was added and heated to 60° C. for 4 hours. The reactionmixture was partitioned between ethyl acetate and water. The combinedorganic phase was dried with MgSO₄, filtered, and concentrated to affordthe The desired product. Then was purified by Gilson HPLC to give pureproduct (10 mg, 3%). LC-MS m/z 541.2. ¹H NMR (DMSO-d₆) δ 1.7 (m, 2H),2.65 (s, 3), 3.09 (m, 3H), 3.27 (m, 3H), 3.7 (m, 2H), 6.33 (d, 1H), 7.21(t, 1H), 7.45 (m, 2H), 7.58 (d, 1H), 7.7 (d, 1H), 8.61 (s, 1H), 10.34(s, 1H).

Example 28 Preparation ofN-(2-Bromophenyl)-N′-[4-chloro-2-hydroxy-3-(piperazine-aminosulfonylphenyl)phenyl]cyanoguanidine

a)4(2-tert-Butyl-6-chloro-benzooxazole-7-sulfonyl)-piperazine-1-carboxylicAcid tert-Butyl Ester

Following the general procedure for the synthesis of sulfonylamidesoutlined in example 51, 2-tert-butyl-6-chlorbenzooxazole-7-sulfonylchloride (5.0 g, 16.2 mmol), triethylamine (2.4 mL, 17.2 mmol), andpiperazine-carboxylic acid tert-butyl ester (3.62 g, 19.4 mmol) werereacted in THF (50 mL) to afford the title compound (5.44 g, 67%). LCMSm/z 402(M−H)⁺ (desired -Boc).

b) 6-Amino3chloro-2-(piperazine-1-sulfonyl)-phenol

Following the general procedure for the hydrolysis of the benzooxazoleto the desired aniline outlined in example 51,4(2-tert-butyl-6-chloro-benzooxazole-7-sulfonyl)-piperazine-1-carboxylicacid tert-butyl ester (2.0 g, 4.3 mmol), water (3.65 mL), and H₂SO₄(3.65 mL) in 1,4-dioxane (60 mL) were reacted to afford the titlecompound (1.22 g, 96%). LCMS m/z 292(M−H)⁺.

c)6-Amino-3-chloro-2-[(4-N-9-fluorenylmethylformate)piperazine-1-sulfonyl)]-phenol

Following the general procedure for protection of amino outlined inexample 51, 6Amino-3-chloro-2-(piperazine-1-sulfonyl)-phenol (660 mg,2.26 mmol), 10% of sodium carbonate (6.0 mL) and 9-fluorenylmethylchloroformate (584.7 mg, 2.26 mmol) in 1,4-dioxane (6.78 mL) werereacted to afford the title compound (1.22 g, crude). LC-MS m/z 514(M⁺).

d)N-(2-Bromophenyl)-N′-[4-chloro-2-hydroxy-3-[(4-N-9-fluorenylmethylformate)piperazineAminosulfonylphenyl]thiourea

Following the general procedure for thiourea formation outlined inexample 12,6-Amino-3-chloro-2-[(4-N-9-fluorenylmethylformate)piperazine-1-sulfonyl)]-phenol(1.22 g, 2.4 mmol, not clean) and 2-bromophenylisothiocyanate (508 mg,2.4 mmol) were coupled to form the desired thiourea (490 mg, 28%). EL-MSm/z 727.07 (M⁻).

e)N-(2-Bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(4-N-9-fluorenylmethylformate)piperazineAminosulfonylphenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 12,N-(2-bromophenyl)-N′-[4-chloro-2-hydroxy-3-[(4-N-9-fluorenylmethylformate)piperazineaminosulfonylphenyl]thiourea (490 mg, 0.67 mmol),tert-butyldimethylsilyl chloride (505 mg, 3.35 mmol) and imidazole (93mg, 1.34 mmol) were reacted to form the desired product (407 mg, 73%).EI-MS m/z 841.08 (M⁺).

f)N-(2-Bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(4-N-9-fluorenylmethylformate)piperazineAminosulfonylphenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 12,N-(2-bromophenyl)-N′-4-chloro-2-tert-butyldimethylsilyloxy-3-[(4-N-9-fluorenylmethylformate)piperazineaminosulfonylphenyl]thiourea (640 mg, 0.76 mmol), methanesulfonylchloride (0.13 mL, 1.52 mmol) and triethylamine (0.25 mL, 1.52 mmol)were reacted. to form the desired product (640 mg, crude). ¹H NMR(CDCl₃) δ 0.4 (s, 6H), 1.1 (s, 9H), 3.2 (m, 4H), 3.48 (m, 4H), 4.22 (t,1H), 4.51 (d, 2H), 7.07-7.44 (m, 9H), 7.54 (d, 2H), 7.6 (d, 1H), 7.74(d, 2H).

g) N-(2-Bromophenyl)-N′-[4-chloro-2-hydroxy-3-(piperazineAminosulfonyl)phenyl]cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 12,N-(2-bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(4-N-9-fluorenylmethylformate)piperazineaminosulfonylphenyl]carbodiimide (640 mg, 0.79 mmol), cyanamide (133 mg,3.16 mmol) and N,N-diisopropylethylamine (122 mg, 0.95 mmol) werereacted, followed by desilylation with Cesium fluoride (144 mg, 0.95mmol) and deprotected amino with 20% piperidine (8 mL) in THF (40 mL) atroom temperature for 30 minutes to form the desired product(purified byGilson HPLC, 346 mg, 76%). LC-MS m/z 513.2. ¹H NMR (DMSO-d₆) δ 3.02 (t,4H), 3.43 (t, 4H), 6.08 (d, 1H), 7.2 (m, 1H), 7.38 (m, 3H), 7.67 (d,1H), 8.84 (s, 1H).

Example 29 Preparation ofN-(2-Chloro-3-fluorophenyl)-N′-[4-chloro-2-hydroxy-3-(piperazine-aminosulfonylpheny)phenyl]cyanoguanidine

a)N-(2-Chloro-3-fluorophenyl)-N′-[4-chloro-2-hydroxy-3-[4-N-9-fluorenylmethylformate)piperazineAminosulfonylphenyl]thiourea

Following the general procedure for thiourea formation outlined inexample 12,6Amino-3-chloro-2-[(4-N-9-fluorenylmethylformate)piperazine-1-sulfonyl)]-phenol(1.0 g, 1.94 mmol, not clean) and 2-chloro-3-fluorophenylisothiocyanate(See Example 45, 400 mg, 1.94 mmol) were coupled to form the desiredthiourea (713 mg, 48%). EL-MS m/z 700.70 (M⁻).

b)N-(2-Chloro-3-fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(4-N-9-fluorenylmethylformate)piperazineAminosulfonylphenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 12,N-(2-chloro-3-fluorophenyl)-N′-[4-chloro-2-hydroxy-3-[(4-N-9-fluorenylmethylformate)piperazineaminosulfonylphenyl]thiourea (713 mg, 1.02 mmol),tert-butyldimethylsilyl chloride (765 mg, 5.1 mmol) and imidazole (139mg, 2.04 mmol) were reacted to form the desired product (455 mg, 55%).EI-MS m/z 814.68 (M⁻).

c)N-(2-Chloro-3-fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(4-N-9-fluorenylmethylformate)piperazineAminosulfonylphenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 12,N-(2-chloro-3-fluorophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(4-N-9-fluorenylmethylformate)piperazineaminosulfonylphenyl]thiourea (455 mg, 0.56 mmol), methanesulfonylchloride (0.1 mL, 1.12 mmol) and triethylamine (0.18 mL, 1.12 mmol) werereacted to form the desired product (537 mg, crude). ¹H NMR (CDCl₃) δ0.39 (s, 6H), 1.04 (s, 9H), 3.19 (m, 4H), 3.49 (m, 4H), 4.23 (t, 1H),4.5 (d, 1H), 6.96 (t, 1H), 7.01-7.41 (m, 9H), 7.55 (d, 1H), 7.74 (d,1H).

d) N-(2-Chloro-3-fluorophenyl)-N′-[4-chloro-2-hydroxy-3-(piperazineAminosulfonyl)phenyl]cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 12,N-(2-chloro-3-fluorophenyl)-N′-[4-chloro-2-tert-butydimethylsilyloxy-3-[(4-N-9-fluorenylmethylformate)piperazineaminosulfonylphenyl]carbodiimide (537 mg, 0.69 mmol), cyanamide (116 mg,2.76 mmol) and N,N-diisopropylethylamine (107 mg, 0.83 mmol) werereacted, followed by desilylation with Cesium fluoride (126 mg, 0.83mmol) and deprotected amino with 20% piperidine (6 mL) in THF (30 mL) atroom temperature for 30 minutes to form the desired product(purified byGilson HPLC, 45 mg, 14%). LC-MS m/z 487.0. ¹H NMR (DMSO-d₆) δ 3.02 (t,4H), 3.43 (t, 4H), 6.2 (d, 1H), 7.2-7.39 (m, 4H), 9.03 (s, 1H).

Example 30 Preparation ofN-(2-Bromophenyl)-N′-[4-chloro-2-hydroxy-3-(4-amino-piperidineAminosulfonylphenyl)phenyl]cyanoguanidine

a)[1-(2tert-Butyl-6-chloro-berizooxazole-7-sulfonyl)-piperidin-4-yl]-carbamicAcid tert-Butyl Ester

Following the general procedure for the synthesis of sulfonylamidesoutlined in example 51, 2-tert-butyl-6-chloro-benzooxazole-7-sulfonylchloride (5.05 g, 16.4 mmol), triethylamine (4.57 mL, 32.8 mmol), and4-N-Boc-aminopiperidine (3.288 g, 16.4 mmol) were reacted in THF (125mL) to afford the title compound (4.18 g, 54%). ¹H NMR (DMSO-d₆) • 7.98(d, 1H, J=8.48 Hz), 7.63 (d, 1H, J=8.47 Hz), 3.73 (d, 2H), 3.35 (bs,2H), 2.92 (m, 2H), 1.75 (d, 2H), 1.35 (s, 10H).

b) 6-Amino-2-(4-amino-piperidine-1-sulfonyl)-3-chloro-phenol

Following the general procedure for the hydrolysis of the benzooxazoleto the desired aniline outlined in example 51,[1-(2-tert-butyl-6-chloro-benzooxazole-7-sulfonyl)-piperidin4-yl]-carbamicacid tert-butyl ester (4.18 g, 8.86 mmol), water (5.5 mL), and H₂SO₄(5.5 mL) in 1,4-dioxane (55mL) were reacted to afford the title compound(2.03 g, 75%). LCMS m/z 306(M−H)⁺.

c)6-Amino-3-chloro-2-[(4-N-9-fluorenylmethylformate)piperidin-1-sulfonyl)]-phenol

Following the general procedure for protection of amino outlined inexample 51, 6Amino-3-chloro-2-(piperidin-1-sulfonyl)-phenol (1.05 g,3.43 mmol), 10% of sodium carbonate (9.1 mL) and 9-fluorenylmethylchloroformate (887.4 mg, 3.43 mmol) in 1,4-dioxane (10.3 mL) werereacted to afford the title compound (1.3 g, crude). LC-MS m/z528.04(M⁺).

d)N-(2-Bromophenyl)-N′-[4-chloro-2-hydroxy-3-[(4-N-9-fluorenylmethylformate)piperidineAminosulfonylphenyl]thiourea

Following the general procedure for thiourea formation outlined inexample 12,6Amino-3-chloro-2-[(4N-9-fluorenylmethylformate)piperidine-1-sulfonyl)]-phenol(1.3 g, 2.42 mmol, not clean) and 2-bromophenylisothiocyanate (520 mg,2.42 mmol) were coupled to form the desired thiourea (700 mg, 39%).EL-MS m/z 739.07 (M⁻).

e)N-(2-Bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(4-N-9-fluorenylmethylformate)piperidineAminosulfonylphenyl]thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 12,N-(2-bromophenyl)-N′-[4-chloro-2-hydroxy-3-[(4-N-9-fluorenylmethylformate)piperazineaminosulfonylphenyl]thiourea (700 mg, 0.95 mmol),tert-butyldimethylsilyl chloride (713 mg, 4.75 mmol) and imidazole (131mg, 1.9 mmol) were reacted to form the desired product (400 mg, 50%).EI-MS m/z 854.52 (M⁻).

f)N-(2-Bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(4-N-9-fluorenylmethylformate)piperidineAminosulfonylphenyl]carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 12,N-(2-bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(4-N-9-fluorenylmethylformate)piperidineaminosulfonylphenyl]thiourea (374 mg, 0.57 mmol), methanesulfonylchloride (0.1 mL, 1.12 mmol) and triethylamine (0.18 mL, 1.12 mmol) werereacted to form the desired product (374 mg, crude). ¹H NMR (CDCl₃) δ0.38 (s, 6H), 1.03 (s, 9H), 1.49 (m, 2H), 1.93 (m, 2H), 2.89 (t, 2H),3.72 (m, 2H), 4.2 (t, 1H), 4.4 (d, 2H), 4.73 (d, 1H), 7.06-7.34 (m, 8H),7.41 (t, 2H), 7.61 (t, 2H), 7.24 (m, 1H), 7.76 (d, 2H).

g) N-(2-Bromophenyl)-N′-[4-chloro-2-hydroxy-3-(piperidineAminosulfonyl)phenyl]cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 12,N-(2-bromophenyl)-N′-[4-chloro-2-tert-butyldimethylsilyloxy-3-[(4-N-9-fluorenylmethylformate)piperidineaminosulfonylphenyl]carbodiimide (400 mg, 0.65 mmol), cyanamide (108 mg,2.6 mmol) and N,N-diisopropylethylamine (99 mg, 0.78 mmol) were reacted,followed by desilylation with Cesium fluoride (123 mg, 0.78 mmol) anddeprotected amino with 20% piperidine (4 mL) in THF (20 mL) at roomtemperature for 30 minutes to form the desired product(purified byGilson HPLC, 105 mg, 41%). LC-MS m/z 527.2. ¹H NMR (DMSO-d₆) δ 1.43 (m,2H), 1.83 (m, 2H), 2.9 (t, 2H), 2.95 (m, 2H), 3.66 (d, 2H), 6.09 (d,1H), 7.14 (t, 1H), 7.34 (m, 3H), 7.64 (d, 1H).

Example 31 Preparation of N-(2-Bromophenyl)-N′-{4-chloro-2-hydroxy-3-[(R)-3-amino-pyrrolidine]aminosulfonylphenyl)phenyl}cyanoguanidine

a)[(R)-1-(2-tert-Butyl-6-chloro-benzooxazole-7-sulfonyl)-pyrrolidin-3-yl]-carbamicAcid tert-Butyl Ester

Following the general procedure for the synthesis of sulfonylamidesoutlined in example 51, 2-tert-butyl-6-chloro-benzooxazole-7-sulfonylchloride (3.0 g, 9.74 mmol), triethylamine (1.63 mL, 11.7 mmol), and(R)-pyrrolidin-3-yl-carbamic acid tert-butyl ester (2.18 g, 11.7 mmol)were reacted in THF (30 mL) to afford the title compound (3.0 g, 67%).¹H NMR (CDCl₃) • 7.77 (d, 1H, J=8.5 Hz), 7.48 (d, 1H, J=8.5 Hz), 4.67(bm, 1H), 4.22 (bm, 1H), 3.66 (bm, 2H), 3.52 (bm, 1H), 3.41 (bm, 1H),2.19 (bm, 1H), 1.90 (bm, 1H), 1.48 (s, 9H).

b) 6-Amino-2-((R)-3-amino-pyrrolidine-1-sulfonyl)-3-chloro-phenol

Following the general procedure for the hydrolysis of the benzooxazoleto the desired aniline outlined in example 51,[(R)-1-(2-tert-butyl-6-chloro-benzooxazole-7-sulfonyl)-pyrrolidin-3-yl]-carbamicacid tert-butyl ester (2.0 g, 4.31 mmol), water (3.6 mL), and H₂SO₄ (3.6mL) in 1,4-dioxane (60 mL) were reacted to afford the title compound(1.2 g, 94%). LCMS m/z 292(M−H)⁺.

c) 6-Amino-3-chloro-2-[(R)-3-N-9-fluorenylmethylformate)pyrrolidine-1-sulfonyl)]-phenol

Following the general procedure for protection of amino outlined inexample 51,6Amino-2-((R)-3-amino-pyrrolidine-1-sulfonyl)-3-chloro-phenol (907 mg,3.1 mmol), 10% of sodium carbonate (7.5 mL) and 9-fluorenylmethylchloroformate(802 mg, 3.1 mmol) in 1,4-dioxane (20 mL) were reacted toafford the title compound (1.7 g, crude). LC-MS m/z 514.2 (M⁺).

d)N-(2-Bromophenyl)-N′-{chloro-2-hydroxy-3-[(R)-3-N-9-fluorenylmethylformate)pyrrolidine]aminosulfonylphenyl}thiourea

Following the general procedure for thiourea formation outlined inexample 12,6-Amino-3-chloro-2-[(R)-3-N-9-fluorenylmethylformate)pyrrolidine-1-sulfonyl)]-phenol(800 mg, 1.56 mmol, not clean) and 2-bromophenylisothiocyanate (333 mg,1.56 mmol) were coupled to form the desired thiourea (405 mg, 36%).EL-MS m/z 727.01 (M⁺).

e)N-(2-Bromophenyl)-N′-{4-chloro-2-tert-butyldimethylsilyloxy-3-[(R)-3-N-9-fluorenylmethylformate)pyrrolidine]aminosulfonylphenyl}thiourea

Following the general procedure for protected phenyl thiourea formationoutlined in example 12,N-(2-bromophenyl)-N′-{4-chloro-2-hydroxy-3-[(R)-3-N-9-fluorenylmethylformate)pyrrolidine]aminosulfonylphenyl}thiourea(405 mg, 0.56 mmol), tert-butyldimethylsilyl chloride (420 mg, 2.8 mmol)and imidazole (76 mg, 1.12 mmol) were reacted to form the desiredproduct (293 mg, 63%). EI-MS m/z 841.2 (M⁺).

f)N-(2-Bromophenyl)-N′-N{4-chloro-2-tert-butyldimethylsilyloxy-3-[(R)-3-N-9-fluorenylmethylformate)pyrrolidine]aminosulfonylphenyl}carbodiimide

Following the general procedure for carbodiimide formation outlined inexample 12,N-(2-bromophenyl)-N′-{chloro-2-tert-butyldimethylsilyloxy-3-[(R)-3-N-9-fluorenylmethylformate)pyrrolidine]aminosulfonylphenyl}thiourea(293 mg, 0.35 mmol), methanesulfonyl chloride (0.05 mL, 0.7 mmol) andtriethylamine (0.1 mL, 0.7 mmol) were reacted to form the desiredproduct (293 mg, crude). ¹H NMR (CDCl₃) δ 0.39 (s, 6H), 1.05 (s, 9H),1.90 (m, 1H), 2.18 (m, 1H), 3.14 (m, 1H), 3.39 (m, 3H), 3.48 (m, 1H),4.2 (t, 1H), 4.4 (d, 2H), 5.06 (d, 1H), 7.16-7.43 (m, 10H), 7.59 (d,1H), 7.7 (d, 1H).

g)N-(2-Bromophenyl)-N′-{4-chloro-2-hydroxy-3-[(R)-3-aminopyrrolidine]aminosulfonylphenyl}cyanoguanidine

Following the general procedure for cyanoguanidine formation outlined inexample 12,N-(2-bromophenyl)-N′-{4-chloro-2-tert-butyldimethylsilyloxy-3-[(R)-3-N-9-fluorenylmethylformate)pyrrolidine]aminosulfonylphenyl}carbodiimide (293 mg, 0.36 mmol),cyanamide (61 mg, 1.44 mmol) and N,N-diisopropylethylamine (56 mg, 0.43mmol) were reacted, followed by desilylation with Cesium fluoride (66mg, 0.43 mmol) and deprotected amino with 20% piperidine (4 mL) in THF(20 mL) at room temperature for 30 minutes to form the desired product(purified by Gilson HPLC, 53 mg, 25%). LC-MS m/z 513.2. ¹H NMR (DMSO-d₆)δ 1.6 (m, 2H), 2.0 (m, 1H), 2.2 (m, 1H), 2.95 (t, 1H), 3.35 (m, 1H),3.50 (m, 2H), 3.72 (m, 1H), 6.9 (d, 1H), 7.2 (t, 1H), 7.42 (m, 2H), 7.7(t, 2H), 8.32 (s, 1H).

METHOD OF TREATMENT

The compounds of Formula (I), or a pharmaceutically acceptable saltthereof can be used in the manufacture of a medicine for theprophylactic or therapeutic treatment of any disease state in a human,or other mammal, which is exacerbated or caused by excessive orunregulated IL-8 cytokine production by such mammal's cell, such as butnot limited to monocytes and/or macrophages, or other chemokines whichbind to the IL 8 α or β receptor, also referred to as the type I or typeII receptor.

Accordingly, the present invention provides a method of treating achemokie mediated disease, wherein the chemokine is one which binds toan IL-8 α or β receptor and which method comprises administering aneffective amount of a compound of Formula (I) or a pharmaceuticallyacceptable salt thereof. In particular, the chemokines are IL-8, GROα,GROβ, GROγ, NAP-2 or ENA-78.

The compounds of Formula (I) are administered in an amount sufficient toinhibit cytokine function, in particular IL8, GROα, GROβ, GROγ, NAP-2 orENA-78, such that they are biologically regulated down to normal levelsof physiological function, or in some case to subnormal levels, so as toameliorate the disease state. Abnormal levels of IL-8, GROα, GROβ, GROγ,NAP-2 or ENA-78 for instance in the context of the present invention,constitute: (i) levels of free IL-8 greater than or equal to 1 picogramper mL; (ii) any cell associated EL-8, GROα, GROβ, GROγ, NAP-2 or ENA-78above normal physiological levels; or (iii) the presence of IL-8, GROα,GROβ, GROγ, NAP-2 or ENA-78 above basal levels in cells or tissues inwhich IL-8, GROα, GROβ, GROγ, NAP-2 or ENA-78 respectively, is produced.

The compounds of Formula (I), in generally have been shown to have alonger t_(½) and improved oral bioavailabilty over the compoundsdisclosed in WO 96/25157 and WO 97/29743 whose disclosures areincorporated herein by reference.

There are many disease states in which excessive or unregulated IL-8production is implicated in exacerbating and/or causing the disease.Chemokine mediated diseases include psoriasis, atopic dermatitis, osteoarthritis, rheumatoid arthritis, asthma, chronic obstructive pulmonarydisease, adult respiratory distress syndrome, inflammatory boweldisease, Crohn's disease, ulcerative colitis, stroke, septic shock,multiple sclerosis, endotoxic shock, gram negative sepsis, toxic shocksyndrome, cardiac and renal reperfusion injury, glomerulonephritis,thrombosis, graft vs. host reaction, Alzheimer's disease, allograftrejections, malaria, restenosis, angiogenesis, atherosclerosis,osteoporosis, gingivitis and undesired hematopoietic stem cells releaseand diseases caused by respiratory viruses, herpesviruses, and hepatitisviruses, meningitis, cystic fibrosis, pre-term labor, cough, pruzitus,multi-organ dysfunction, trauma, strains, sprains, contusions, psoriaticarthritis, herpes, encephalitis, CNS vasculitis, traumatic brain injury,CNS tumors, subarachnoid hemorrhage, post surgical trauma, interstitialpneumonitis, hypersensitivity, crystal induced arthritis, acute andchronic pancreatitis, acute alcoholic hepatitis, necrotizingenterocolitis, chronic sinusitis, uveitis, polymyositis, vasculitis,acne, gastric and duodenal ulcers, celiac disease, esophagitis,glossitis, airflow obstruction, airway hyperresponsiveness,bronchiolitis obliterans organizing pneumonia, bronchiectasis,bronchiolitis, bronchiolitis obliterans, chronic bronchitis, corpulmonae, dyspnea, emphysema, hypercapnea, hyperinflation, hypoxemia,hyperoxia-induced inflammations, hypoxia, surgical lung volumereduction, pulmonary fibrosis, pulmonary hypertension, right ventricularhypertropy, sarcoidosis, small airway disease, ventilation-perfusionmismatching, wheeze, colds and lupus.

These diseases are primarily characterized by massive neutrophilinfiltration, T-cell infiltration, or neovascular growth, and areassociated with increased IL-8, GROα, GROβ, GROγ, NAP-2 or ENA-78production which is responsible for the chemotaxis of neutrophils intothe inflammatory site or the directional growth of endothelial cells. Incontrast to other inflammatory cytokines (IL-1, TNF, and IL-6), I8,GROα, GROβ, GROγ, NAP-2 or ENA-78 have the unique property of promotingneutrophil chemotaxis, enzyme release including but not limited toelastase release as well as superoxide production and activation. Thecc-chemokines but particularly, GROα, GROβ, GROγ, NAP-2 or ENA-78,working through the IL-8 type I or II receptor can promote theneovascularization of tumors by promoting the directional growth ofendothelial cells. Therefore, the inhibition of IL-8 induced chemotaxisor activation would lead to a direct reduction in the neutrophilinfiltration.

Recent evidence also implicates the role of chemokines in the treatmentof HIV infections, Littleman et al., Nature 381, pp. 661 (1996) and Koupet al., Nature 381, pp. 667 (1996).

Present evidence also indicates the use of IL-8 inhibitors in thetreatment of atherosclerosis. The first reference, Boisvert et al., J.Clin. Invest, 1998, 101:353-363 shows, through bone marrowtransplantation, that the absence of IL-8 receptors on stem cells (and,therefore, on monocytes/macrophages) leads to a reduction in thedevelopment of atherosclerotic plaques in LDL receptor deficient mice.Additional supporting references are: Apostolopoulos, et al.,Arterioscler. Thromb. Vasc. Biol. 1996, 16:1007-1012; Liu, et al.,Arterioscler. Thromb. Vasc. Biol, 1997, 17:317-323; Rus, et al.,Atherosclerosis. 1996, 127:263-271.; Wang et al., J. Biol. Chem. 1996,271:8837-8842; Yue, et al., Eur. J. Pharmacol. 1993, 240:81-84; Koch, etal., Am. J. Pathol., 1993, 142:1423-1431.; Lee, et al., Immunol. Lett.,1996, 53, 109-113.; and Terkeltaub et al., Arterioscler. Thromb., 1994,14:47-53.

The present invention also provides for a means of treating, in an acutesetting, as well as preventing, in those individuals deemed susceptibleto, CNS injuries by the chemokine receptor antagonist compounds ofFormula (I).

CNS injuries as defined herein include both open or penetrating headtrauma, such as by surgery, or a closed head trauma injury, such as byan injury to the head region. Also included within this definition isischemic stroke, particularly to the brain area.

Ischemic stroke may be defined as a focal neurologic disorder thatresults from insufficient blood supply to a particular brain area,usually as a consequence of an embolus, thrombi, or local atheromatousclosure of the blood vessel. The role of inflammatory cytokines in thisarea has been emerging and the present invention provides a mean for thepotential treatment of these injuries. Relatively little treatment, foran acute injury such as these has been available.

TNF-α is a cytokine with proinflammatory actions, including endothelialleukocyte adhesion molecule expression. Leukocytes infiltrate intoischemic brain lesions and hence compounds, which inhibit or decreaselevels of TNF would be useful for treatment of ischemic brain injury.See Liu et al., Stroke, Vol. 25., No. 7, pp. 1481-88 (1994) whosedisclosure is incorporated herein by reference.

Models of closed head injuries and treatment with mixed 5-LO/CO agentsis discussed in Shohami et al., J. of Vaisc & Clinical Physiology andPharmacology, Vol. 3, No. 2, pp. 99-107 (1992) whose disclosure isincorporated herein by reference. Treatment, which reduced edemaformation, was found to improve functional outcome in those animalstreated.

The compounds of Formula (I) are administered in an amount sufficient toinhibit IL8, binding to the IL-8 alpha or beta receptors, from bindingto these receptors, such as evidenced by a reduction in neutrophilchemotaxis and activation. The discovery that the compounds of Formula(I) are inhibitors of IL-8 binding is based upon the effects of thecompounds of Formulas (I) in the in vitro receptor binding assays whichare described herein. The compounds of Formula (I) have been shown to beinhibitors of type II IL-8 receptors.

As used herein, the term “IL-8 mediated disease or disease state” refersto any and all disease states in which IL-8, GROα, GROβ, GROγ, NAP-2 orENA-78 plays a role, either by production of IL-8, GROα, GROβ, GROγ,NAP-2 or ENA-78 themselves, or by IL,8, GROα, GROβ, GROγ, NAP-2 orENA-78 causing another monokine to be released, such as but not limitedto IL-1, IL-6 or TNF. A disease state in which, for instance, IL-1 is amajor component, and whose production or action, is exacerbated orsecreted in response to IL-8, would therefore be considered a diseasestate mediated by IL-8.

As used herein, the term “chemokine mediated disease or disease state”refers to any and all disease states in which a chemokine which binds toan IL-8 α or β receptor plays a role, such as but not limited to IL-8,GRO-α, GRO-β, GROγ, NAP-2 or ENA-78. This would include a disease statein which, IL-8 plays a role, either by production of IL-8 itself, or byIL-8 causing another monokine to be released, such as but not limited toIL-1, IL-6 or TNF. A disease state in which, for instance, IL-1 is amajor component, and whose production or action, is exacerbated orsecreted in response to IL-8, would therefore be considered a diseasestated mediated by IL-8.

As used herein, the term “cytokine” refers to any secreted polypeptidethat affects the functions of cells and is a molecule, which modulatesinteractions between cells in the immune, inflammatory or hematopoieticresponse. A cytokine includes, but is not limited to, monokines andlymphokines, regardless of which cells produce them. For instance, amonokine is generally referred to as being produced and secreted by amononuclear cell, such as a macrophage and/or monocyte. Many other cellshowever also produce monokines, such as natural killer cells,fibroblasts, basophils, neutrophils, endothelial cells, brainastrocytes, bone marrow stromal cells, epideral keratinocytes andB-lymphocytes. Lymphokines are generally referred to as being producedby lymphocyte cells. Examples of cytokines include, but are not limitedto, Interleukin-1 (IL1), Interleukin-6 (IL6), Interleukin-8 (IL-8),Tumor Necrosis Factor-alpha (TNF-α) and Tumor Necrosis Factor beta(TNF-β).

As used herein, the term “chemokine” refers to any secreted polypeptidethat affects the functions of cells and is a molecule which modulatesinteractions between cells in the immune, inflammatory or hematopoieticresponse, similar to the term “cytokine” above. A chemokine is primarilysecreted through cell transmembranes and causes chemotaxis andactivation of specific white blood cells and leukocytes, neutrophils,monocytes, macrophages, T-cells, B-cells, endothelial cells and smoothmuscle cells. Examples of chemokines include, but are not limited toIL-8, GRO-α, GRO-β, GRO-γ, NAP-2, ENA-78, IP-10, MIP-1α, MIP-β, PF4, andMCP 1, 2, and 3.

The present compounds are also useful in normalizing leukocyte counts aswell as normalizing levels of circulating chemokines.

In order to use a compound of Formula (I) or a pharmaceuticallyacceptable salt thereof in therapy, it will normally be formulated intoa pharmaceutical composition in accordance with standard pharmaceuticalpractice. This invention, therefore, also relates to a pharmaceuticalcomposition comprising an effective, non-toxic amount of a compound ofFormula (I) and a pharmaceutically acceptable carrier or diluent.

Compounds of Formula (I), pharmaceutically acceptable salts thereof andpharmaceutical compositions incorporating such may conveniently beadministered by any of the routes conventionally used for drugadministration, for instance, orally, topically, parenterally or byinhalation. The compounds of Formula (I) may be administered inconventional dosage forms prepared by combining a compound of Formula(I) with standard pharmaceutical carriers according to conventionalprocedures. The compounds of Formula (I) may also be administered inconventional dosages in combination with a known, second therapeuticallyactive compound. These procedures may involve mixing, granulating andcompressing or dissolving the ingredients as appropriate to the desiredpreparation. It will be appreciated that the form and character of thepharmaceutically acceptable character or diluent is dictated by theamount of active ingredient with which it is to be combined, the routeof administration and other well-known variables. The carrier(s) must be“acceptable” in the sense of being compatible with the other ingredientsof the formulation and not deleterious to the recipient thereof.

The pharmaceutical carrier employed may be, for example, either a solidor liquid. Exemplary of solid carriers are lactose, terra alba, sucrose,talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acidand the like. Exemplary of liquid carriers are syrup, peanut oil, oliveoil, water and the like. Similarly, the carrier or diluent may includetime delay material well known to the art, such as glycerylmono-stearate or glyceryl distearate alone or with a wax.

A wide variety of pharmaceutical forms can be employed. Thus, if a solidcarrier is used, the preparation can be tableted, placed in a hardgelatin capsule in powder or pellet form or in the form of a troche orlozenge. The amount of solid carrier will vary widely but preferablywill be from about 25 mg to about 1 g. When a liquid carrier is used,the preparation will be in the form of a syrup, emulsion, soft gelatincapsule, sterile injectable liquid such as an ampule or nonaqueousliquid suspension.

Compounds of Formula (I) may be administered topically, that is bynon-systemic administration. This includes the application of a compoundof Formula (I) externally to the epidermis or the buccal cavity and theinstillation of such a compound into the ear, eye and nose, such thatthe compound does not significantly enter the blood stream. In contrast,systemic administration refers to oral, intravenous, intraperitoneal andintramuscular administration.

Formulations suitable for topical administration include liquid orsemi-liquid preparations suitable for penetration through the skin tothe site of inflammation such as liniments, lotions, creams, ointmentsor pastes, and drops suitable for administration to the eye, ear ornose. The active ingredient may comprise, for topical administration,from 0.001% to 10% w/w, for instance from 1% to 2% by weight of theFormulation. It may however comprise as much as 10% w/w but preferablywill comprise less than 5% w/w, more preferably from 0.1% to 1% w/w ofthe Formulation.

Lotions according to the present invention include those suitable forapplication to the skin or eye. An eye lotion may comprise a sterileaqueous solution optionally containing a bactericide and may be preparedby methods similar to those for the preparation of drops. Lotions orliniments for application to the skin may also include an agent tohasten drying and to cool the skin, such as an alcohol or acetone,and/or a moisturizer such as glycerol or an oil such as castor oil orarachis oil.

Creams, ointments or pastes according to the present invention aresemi-solid formulations of the active ingredient for externalapplication. They may be made by mixing the active ingredient in finelydivided or powdered form, alone or in solution or suspension in anaqueous or non-aqueous fluid, with the aid of suitable machinery, with agreasy or non-greasy base. The base may comprise hydrocarbons such ashard, soft or liquid paraffin, glycerol, beeswax, a metallic soap; amucilage; an oil of natural origin such as almond, corn, arachis, castoror olive oil; wool fat or its derivatives or a fatty acid such as stericor oleic acid together with an alcohol such as propylene glycol or amacrogel. The formulation may incorporate any suitable surface activeagent such as an anionic, cationic or non-ionic surfactant such as asorbitan ester or a polyoxyethylene derivative thereof. Suspendingagents such as natural gums, cellulose derivatives or inorganicmaterials such as silicaceous silicas, and other ingredients such aslanolin, may also be included.

Drops according to the present invention may comprise sterile aqueous oroily solutions or suspensions and may be prepared by dissolving theactive ingredient in a suitable aqueous solution of a bactericidaland/or fungicidal agent and/or any other suitable preservative, andpreferably including a surface active agent. The resulting solution maythen be clarified by filtration, transferred to a suitable containerwhich is then sealed and sterilized by autoclaving or maintaining at98-100° C. for half an hour. Alternatively, the solution may besterilized by filtration and transferred to the container by an aseptictechnique. Examples of bactericidal and fungicidal agents suitable forinclusion in the drops are phenylmercuric nitrate or acetate (0.002%),benzalkonium chloride (0.01%) and chlorhexidine acetate (0.01%).Suitable solvents for the preparation of an oily solution includeglycerol, diluted alcohol and propylene glycol.

Compounds of formula (I) may be administered parenterally, that is byintravenous, intramuscular, subcutaneous intranasal, intrarectal,intravaginal or intraperitoneal administration. The subcutaneous andintramuscular forms of parenteral administration are generallypreferred. Appropriate dosage forms for such administration may beprepared by conventional techniques. Compounds of Formula (I) may alsobe administered by inhalation that is by intranasal and oral inhalationadministration. Appropriate dosage forms for such administration, suchas an aerosol formulation or a metered dose inhaler, may be prepared byconventional techniques.

For all methods of use disclosed herein for the compounds of Formula (I)the daily oral dosage regimen will preferably be from about 0.01 toabout 80 mg/kg of total body weight. The daily parenteral dosage regimenabout 0.001 to about 80 mg/kg of total body weight. The daily topicaldosage regimen will preferably be from 0.1 mg to 150 mg, administeredone to four, preferably two or three times daily. The daily inhalationdosage regimen will preferably be from about 0.01 mg/kg to about I mg/kgper day. It will also be recognized by one of skill in the art that theoptimal quantity and spacing of individual dosages of a compound ofFormula (I) or a pharmaceutically acceptable salt thereof will bedetermined by the nature and extent of the condition being treated, theform, route and site of administration, and the particular patient beingtreated, and that such optimums can be determined by conventionaltechniques. It will also be appreciated by one of skill in the art thatthe optimal course of treatment, i.e., the number of doses of a compoundof Formula (I) or a pharmaceutically acceptable salt thereof given perday for a defined number of days, can be ascertained by those skilled inthe art using conventional course of treatment determination tests.

The invention will now be described by reference to the followingbiological examples which are merely illustrative and are not to beconstrued as a limitation of the scope of the present invention.

BIOLOGICAL EXAMPLES

The IL-8, and GRO-α chemokine inhibitory effects of compounds of thepresent invention are determined by the following in vitro assay:

Receptor Binding Assays

[¹²⁵I] IL-8 (human recombinant) is obtained from Amersham Corp.,Arlington Heights, Ill., with specific activity 2000 Ci/mmol. GRO-α isobtained from NEN-New England Nuclear. All other chemicals are ofanalytical grade. High levels of recombinant human IL-8 type α and βreceptors were individually expressed in Chinese hamster ovary cells asdescribed previously (Holmes, et al., Science, 1991, 253, 1278). TheChinese hamster ovary membranes were homogenized according to apreviously described protocol (Haour, et al., J. Biol. Chem., 249 pp2195-2205 (1974)). Except that the homogenization buffer is changed to10 mM Tris-HCL, 1 mM MgSO₄, 0.5 mM EDTA (ethylene-diaminetetra-aceticacid), 1 mM PMSF (α-toluenesulphonyl fluoride), 0.5 mg/L Leupeptin, pH7.5. Membrane protein concentration is determined using Pierce Co.micro-assay kit using bovine serum albumin as a standard. All assays areperformed in a 96-well micro plate format. Each reaction mixturecontains ¹²⁵I IL-8 (0.25 nM) or ¹²⁵I GRO-α and 0.5 μg/mL of IL-8Rα or1.0 μg/mL of IL-8Rβ membranes in 20 mM Bis-Trispropane and 0.4 mM TrisHCl buffers, pH 8.0, containing 1.2 mM MgSO₄, 0.1 mM EDTA, 25 mM Na and0.03% CHAPS. In addition, drug or compound of interest is added whichhas been pre-dissolved in DMSO so as to reach a final concentration ofbetween 0.01 nM and 100 uM. The assay is initiated by addition of¹²⁵I-IL8. After 1 hour at room temperature the plate is harvested usinga Tomtec 96-well harvester onto a glass fiber filtermat blocked with 1%polyethylenimine/0.5% BSA and washed 3 times with 25 mM NaCl, 10 mMTrisHCl, 1 mM MgSO₄, 0.5 mM EDTA, 0.03% CHAPS, pH 7.4. The filter isthen dried and counted on the Betaplate liquid scintillation counter.The recombinant IL-8 Rα, or Type I, receptor is also referred to hereinas the non-permissive receptor and the recombinant IL-8 Rβ, or Type II,receptor is referred to as the permissive receptor.

Representative compounds of Formula (I), Examples 1 to 106 haveexhibited positive inhibitory activity in this assay at IC₅₀ levels <30uM.

Chemotaxis Assay

The in vitro inhibitory properties of these compounds are determined inthe neutrophil chemotaxis assay as described in Current Protocols inImmunology, vol. I, Suppl 1, Unit 6.12.3., whose disclosure isincorporated herein by reference in its entirety. Neutrophils whereisolated from human blood as described in Current Protocols inImmunology Vol. I, Suppl 1 Unit 7.23.1, whose disclosure is incorporatedherein by reference in its entirety. The chemoattractants IL8, GRO-α,GRO-β, GRO-γ and NAP-2 are placed in the bottom chamber of a 48multiwell chamber (Neuro Probe, Cabin John, Md.) at a concentrationbetween 0.1 and 100 nM. The two chambers are separated by a 5 uMpolycarbonate filter. When compounds of this invention are tested, theyare mixed with the cells (0.001-1000 nM) just prior to the addition ofthe cells to the upper chamber. Incubation is allowed to proceed forbetween about 45 and 90 min at about 37° C. in a humidified incubatorwith 5% CO₂. At the end of the incubation period, the polycarbonatemembrane is removed and the topside washed, the membrane then stainedusing the Diff Quick staining protocol (Baxter Products, McGaw Park,Ill., USA). Cells which have chemotaxed to the chemokine are visuallycounted using a microscope. Generally, four fields are counted for eachsample, these numbers are averaged to give the average number of cellswhich had migrated. Each sample is tested in triplicate and eachcompound repeated at least four times. To certain cells (positivecontrol cells) no compound is added, these cells represent the maximumchemotactic response of the cells. In the case where a negative control(unstimulated) is desired, no chemokine is added to the bottom chamber.The difference between the positive control and the negative controlrepresents the chemotactic activity of the cells.

Elastase Release Assay

The compounds of this invention are tested for their ability to preventElastase release from human neutrophils. Neutrophils are isolated fromhuman blood as described in Current Protocols in Immunology Vol. I,Suppl 1 Unit 7.23.1. PMNs 0.88×10⁶ cells suspended in Ringer's Solution(NaCl 118, KCl 4.56, NaHCO₃ 25, KH₂PO₄ 1.03, Glucose 11.1, HEPES 5 mM,pH 7.4) are placed in each well of a 96 well plate in a volume of 50 ul.To this plate is added the test compound (0.001-1000 nM) in a volume of50 ul, Cytochalasin B in a volume of 50 ul (20 ug/ml) and Ringers bufferin a volume of 50 ul. These cells are allowed to warm (37° C., 5% CO2,95% RH) for 5 min before IL-8, GROα, GROβ, GROγ or NAP-2 at a finalconcentration of 0.01-1000 nM was added. The reaction is allowed toproceed for 45 min before the 96 well plate is centrifuged (800×g 5min.) and 100 ul of the supernatant removed. This supernatant is addedto a second 96 well plate followed by an artificial elastase substrate(MeOSuc-Ala-Ala-Pro-Val-AMC, Nova Biochem, La Jolla, Calif.) to a finalconcentration of 6 ug/ml dissolved in phosphate buffered saline.Immediately, the plate is placed in a fluorescent 96 well plate reader(Cytofluor 2350, Millipore, Bedford, Mass.) and data collected at 3 minintervals according to the method of Nakajima et al J. Biol. Chem. 2544027 (1979). The amount of Elastase released from the PMNs is calculatedby measuring the rate of MeOSuc-Ala-Ala-Pro-Val-AMC degradation.

TNF-α in Traumatic Brain Injury Assay

The present assay provides for examination of the expression of tumornecrosis factor mRNA in specific brain regions, which followexperimentally, induced lateral fluid-percussion traumatic brain injury(TBI) in rats. Adult Sprague-Dawley rats (n=42) were anesthetized withsodium pentobarbital (60 mg/kg, i.p.) and subjected to lateralfluid-percussion brain injury of moderate severity (2.4 atm.)centeredover the left temporaparietal cortex (n=18), or “sham” treatment(anesthesia and surgery without injury, n=18). Animals are sacrificed bydecapitation at 1, 6 and 24 hr. post injury, brains removed, and tissuesamples of left (injured) parietal cortex (LC), corresponding area inthe contralateral right cortex (RC), cortex adjacent to injured parietalcortex (LA), corresponding adjacent area in the right cortex (RA), lefthippocampus (LH) and right hippocampus (RH) are prepared. Total RNA areisolated and Northern blot hybridization is performed and quantitatedrelative to an TNF-α positive control RNA (macrophage=100%). A markedincrease of 1NF-α mRNA expression is observed in LH (104±17% of positivecontrol, p <0.05 compared with sham), LC (105±21%, p<0.05) and LA(69±8%, p<0.01) in the traumatized hemisphere 1 hr. following injury. Anincreased TNF-α mRNA expression is also observed in LH (46±8%, p<0.05),LC (30±3%, p<0.01) and LA (32±3%, p<0.01) at 6 hr which resolves by 24hr following injury. In the contralateral hemisphere, expression ofTNF-α mRNA is increased in RH (46±2%, p<0.01), RC (4±3%) and RA (22±8%)at 1 hr and in RH (28±11%), RC (7±5%) and RA (26±6%, p<0.05) at 6 hr butnot at 24 hr following injury. In sham (surgery without injury) or naiveanimals, no consistent changes in expression of TNF-α mRNA are observedin any of the 6 brain areas in either hemisphere at any times. Theseresults indicate that following parasagittal fluid-percussion braininjury, the temporal expression of TNF-α mRNA is altered in specificbrain regions, including those of the non-traumatized hemisphere. SinceTNF-α is able to induce nerve growth factor (NGF) and stimulate therelease of other cytokines from activated astrocytes, thispost-traumatic alteration in gene expression of TNF-α plays an importantrole in both the acute and regenerative response to CNS trauma.

CNS Injury Model for IL-1β mRNA

This assay characterizes the regional expression of interleukin-1β(IL-1β) mRNA in specific brain regions following experimental lateralfluid-percussion traumatic brain injury (TBI) in rats. AdultSprague-Dawley rats (n=42) are anesthetized with sodium pentobarbital(60 mg/kg, i.p.) and subjected to lateral fluid-percussion brain injuryof moderate severity (2.4 atm.)centered over the left temporaparietalcortex (n=18), or “sham” treatment (anesthesia and surgery withoutinjury). Animals are sacrificed at 1, 6 and 24 hr. post injury, brainsremoved, and tissue samples of left (injured) parietal cortex (LC),corresponding area in the contralateral right cortex (RC), cortexadjacent to injured parietal cortex (LA), corresponding adjacent area inthe right cortex (RA), left hippocampus (LH) and right hippocampus (RH)are prepared. Total RNA is isolated and Northern blot hybridization wasperformed and the quantity of brain tissue IL-1β mRNA is presented aspercent relative radioactivity of IL-1β positive macrophage RNA whichwas loaded on the same gel. At 1 hr following brain injury, a marked andsignificant increase in expression of IL-1β mRNA is observed in LC(20.0±0.7% of positive control, n=6, p<0.05 compared with sham animal),LH (24.5±0.9%, p<0.05) and LA (21.5±3.1%, p<0.05) in the injuredhemisphere, which remained elevated up to 6 hr. post injury in the LC(4.0±0.4%, n=6, p<0.05) and LH (5.0±1.3%, p<0.05). In sham or naiveanimals, no expression of IL-1β mRNA is observed in any of therespective brain areas. These results indicate that following TBI, thetemporal expression of IL-1β mRNA is regionally stimulated in specificbrain regions. These regional changes in cytokines, such as IL-1β play arole in the post-traumatic.

All publications, including but not limited to patents and patentapplications, cited in this specification are herein incorporated byreference as if each individual publication were specifically andindividually indicated to be incorporated by reference herein as thoughfully set forth.

The above description fully discloses the invention including preferredembodiments thereof. Modifications and improvements of the embodimentsspecifically disclosed herein are within the scope of the followingclaims. Without further elaboration, it is believed that one skilled inthe art can, using the preceding description, utilize the presentinvention to its fullest extent. Therefore the Examples herein are to beconstrued as merely illustrative and not a limitation of the scope ofthe present invention in any way. The embodiments of the invention inwhich an exclusive property or privilege is claimed are defined asfollows.

What is claimed is:
 1. A compound of the formula:

wherein: R is selected from the group consisting of cyano, OR₁₁,C(O)NR₁₅R₁₆, R₁₈, C(O)OR₁₁, C(O)R₁₁, and S(O)₂R₁₇; R′R″ is independentlyselected from the group consisting of hydrogen, NR₆R₇, OH, OR_(a),C₁₋₅alkyl, aryl, arylC₁₋₄alkyl, aryl C2-4alkenyl; cycloalkyl, cycloalkylC₁₋₅ alkyl, heteroaryl, heteroarylC₁₋₄alkyl, heteroarylC₂₋₄ alkenyl,heterocyclic, heterocyclic C₁₋₄alkyl, and a heterocyclic C₂₋₄alkenylmoiety, all of which moieties may be optionally substituted one to threetimes independently by a substituent selected from the group consistingof halogen, nitro, halosubstituted C₁₋₄alkyl, C₁₂₋₄alkyl, amino, mono-or di-C₁₋₄ alkyl substituted amine, OR_(a), C(O)R_(a), NR_(a)C(O)OR_(a),OC(O)NR₆R₇, hydroxy, NR₉C(O)R_(a), S(O)_(m′)R_(a), C(O)NR₆R₇, C(O)OH,C(O)OR_(a), S(O)_(t)NR₆R₇, and NHS(O)_(t)R_(a); or the two R_(b)substituents join to form a 3-10 membered ring, optionally substitutedand containing, in addition to optionally substituted C₁₋₄ alkyl,independently, 1 to 3 NR_(a), O, S, SO, or SO₂ moieties, which moietiescan be optionally unsaturated; R′″ is selected from the group consistingof Y hydrogen, halogen, nitro, cyano, halosubstituted C₁₋₁₀ alkyl, C₁₋₁₀alkyl, C₂₋₁₀ alkenyl, C₁₋₁₀ alkoxy, halosubstituted C₁₋₁₀ alkoxy, azide,(CR₈R₈)_(q)S(O)_(t)R_(a), (CR₈R₈)_(q)OR_(a), hydroxy, hydroxysubstituted C₁₋₄alkyl, aryl, aryl C₁₋₄ alkyl, aryloxy, arylC₁₋₄alkyloxy, aryl C₂₋₁₀ alkenyl, heteroaryl, heteroarylalkyl, heteroarylC₁₋₄ alkyloxy, heteroaryl C₂₋₁₀ alkenyl, heterocyclic, heterocyclicC₁₋₄alkyl, heterocyclicC₂₋₁₀ alkenyl, (CR₈R₈)_(q)NR₄R₅, C₂₋₁₀ alkenylC(O)NR₄R₅, (CR₈R₈)_(q)C(O)NR₄R₅, (CR₈R₈)_(q)C(O)NR₄R₁₀, S(O)₃R₈,(CR₈R₈)_(q)C(O)R₁₁, C₂₋₁₀ alkenylC(O)R₁₁, (CR₈R₈)_(q)C(O)OR₁₁,C₂₋₁₀alkenylC(O)OR₁₁, (CR₈R₈)_(q)OC(O)R₁₁, (CR₈R₈)_(q)NR₄C(O)R₁₁,(CR₈R₈)_(q)NHS(O)_(t)R₁₃, (CR₈R₈)_(q)S(O)_(t)NR₄R₅,(CR₈R₈)_(q)C(NR₄)NR₄R₅, and (CR₈R₈)_(q)NR₄C(NR₅)R₁₁; or two Y moietiestogether form O—(CH₂)_(s)—O or a 5 to 6 membered saturated orunsaturated ring, such that the alkyl, aryl, arylalcyl, heteroaryl,heteroaryl alkyl, heterocyclic, and heterocyclicalkyl groups may beoptionally substituted; R₁ is independently selected from the groupconsisting of hydrogen, halogen, nitro, cyano, C₁₋₁₀ alkyl,halosubstituted C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₁₋₁₀ alkoxy,halosubstituted C₁₋₁₀alkoxy, azide, S(O)_(t)R₄, (CR₈R₈)_(q)S(O)_(t)R₄,hydroxy, hydroxy substituted C₁₋₄alkyl, aryl, aryl C₁₋₄ alkyl, arylC₂₋₁₀ alkenyl, aryloxy, aryl C₁₋₄ alkyloxy, heteroaryl, heteroarylalkyl,heteroaryl C₂₋₁₀ alkenyl, heteroaryl C₁₋₄ alkyloxy, heterocyclic,heterocyclic C₁₋₄alkyl, heterocyclicC₁₋₄alkyloxy, heterocyclicC₂₋₁₀alkenyl, (CR₈R₈)_(q)NR₄R₅, (CR₈R₈)_(q)C(O)NR₄R₅, C₂₋₁₀ alkenylC(O)NR₄R₅, (CR₈R₈)_(q)C(O)NR₄R₁₀, S(O)₃R₈, (CR₈R₈)_(q)C(O)R₁₁, C₂₋₁₀alkenyl C(O)R₁₁, C₂₋₁₀ alkenyl C(O)OR₁₁, (CR₈R₈)_(q)C(O)OR₁₁,(CR₈R₈)_(q)OC(O)R₁₁, (CR₈R₈)_(q)NR₄C(O)R₁₁, (CR₈R₈)_(q)C(NR₄)NR₄R₅,(CR₈R₈)_(q)NR₄C(NR₅)R₁₁, (CR₈R₈)_(q)NHS(O)_(t)R₁₃, and(CR₈R₈)_(q)S(O)_(t)NR₄R₅; or two R₁ moieties together form O—(CH₂)_(s)Oor a 5 to 6 membered saturated or unsaturated ring, wherein the alkyl,aryl, arylalkyl, heteroaryl, and heterocyclic moieties may be optionallysubstituted.
 2. The compound according to claim 1 wherein R₁ issubstituted in the 4-position by an electron withdrawing moiety.
 3. Thecompound according to claim 2 wherein R₁ is halogen, cyano or nitro. 4.The compound according to claim 3 wherein R₁ is halogen.
 5. The compoundaccording to claim 4 wherein R₁ is independently fluorine, chlorine, orbromine.
 6. The compound according to claim 1 wherein Y ismono-substituted in the 2′-position or 3′-position, or is disubstitutedin the 2′- or 3′-position of a monocyclic ring.
 7. The compoundaccording to claim 6 wherein Y is halogen.
 8. The compound according toclaim 4 wherein Y is independently fluorine, chlorine, or bromine. 9.The compound according to claim 1 wherein R_(b) is hydrogen, C₁₋₄ alkyl,C₁₋₄ alkyl substituted with C(O)OH, or C(O)OR_(a).
 10. The compoundaccording to claim 1 wherein Y is halogen, n is 1 or 2, R₁ is halogen, mis 1 or 2, and R_(b) is independently hydrogen, C₁₋₄ alkyl, C₁₋₄ alkylsubstituted with C(O)OH, or C(O)OR_(a).
 11. The compound according toclaim 1 which is selected from the group consisting of:N-(2-bromophenyl)-N′-[4-chloro-2-hydroxy-3-(N″,N″dimethylaminosulfonyl)phenyl]cyanoguanidine;N-[4-chloro-2-hydroxy-3-(N″,N″-dimethylaminosulfonyl)phenyl]-N′-(2,3-dichlorophenyl)cyanoguanidine;N-(2-bromophenyl)-N′-[-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]cyanoguanidine;N-(2,3-dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]cyanoguanidine;N-phenyl-N′-[4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]cyanoguanidine;N-(2-bromophenyl)-N′-[4-chloro-2-hydroxy-3-[R-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]cyanoguanidine;N-(2,3-dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-[R-(2-methoxymethyl)pyrrolidin-1-yl]aminosulfonylphenyl]cyanoguanidine;N-(2-bromophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-isoxazolidinylaminosulfonylphenyl]cyanoguanidine;N-(2,3-dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-isoxazolidinylaninosulfonylphenyl]cyanoguanidine;N-(2-bromophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-tetrahydroisoxazylaminosulfonyl)phenyl]cyanoguanidine;N-(2,3-dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-tetrahydroisoxazylaminosulfonyl)phenyl]cyanoguanidine;N-(2,3-dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-(4-thiomorpholinylaminosulfonyl)phenyl]cyanoguanidine;N-[4-chloro-2-hydroxy-3-[N″,N″-dimethylaminosulfonyl]phenyl]-N′-(2-bromophenyl)propylguanidine;N-(2-bromophenyl)-N′-[4-chloro-2-hydroxy-3-(4oxidothiomorpholino)aminosulfonylphenyl]cyanoguanidine;N-(2,3-chlorophenyl)-N′-[4-chloro-2-hydroxy-3-(4oxidothiomorpholino)aminosulfonylphenyl]cyanoguanidine;N-(2-bromophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-methylpiperazino)aminosulfonylphenyl]cyanoguanidine;N-(2,3-dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-methylpiperazino)aminosulfonylphenyl]cyanoguanidine;N-(2-bromophenyl)-N′-N[4-chloro-2-hydroxy-3-(N″-ethylmorpholino)aminosulfonylphenyl]cyanoguanidine;N-(2,3-dichlorophenyl)-N′-[4-chloro-2-hydroxy-3-(N″-ethylmorpholino)aminosulfonylphenyl]cyanoguanidine;N-(2-bromophenyl)-N′-{4-chloro-2-hydroxy-3-[N″-ethyl-2-(2-ethylpyrrolidino)]aminosulfonylpheny}cyanoguanidine;N-(2,3-dichlorophenyl)-N′-{4-chloro-2-hydroxy-3-[N″-ethyl-2-(2-ethylpyrrolidino)]aminosulfonylpheny}cyanoguanidine;N-(2-bromophenyl)-N′-{4-chloro-2-hydroxy-3-[S-(+)-(2-carboxy)pyrrolidin-1-yl]aminosulfonylpheny}cyanoguanidine;N-(2,3-dichlorophenyl)-N′-{4-chloro-2-hydroxy-3-[S-(+)-(2-carboxy)pyrrolidin-1-yl]aminosulfonylpheny}cyanoguanidine;N-(2-bromo-3-fluorophenyl)-N′-[4-chloro2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]sulfonylphenyl]cyanoguanidine;N-(2-phenoxyphenyl)-N′-[4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]sulfonylphenyl]cyanoguanidine;andN-(2-benzoxyphenyl)-N′-[4-chloro-2-hydroxy-3-[S-(+)-(2-methoxymethyl)pyrrolidin-1-yl]sulfonylphenyl]cyanoguanidine;or a pharmaceutically acceptable salt thereof.
 12. A pharmaceuticalcomposition comprising a compound according to claim 1 and apharmaceutically acceptable carrier or diluent.
 13. A method of treatinga chemokine mediated disease, wherein the chemokine binds to an IL-8 aor b receptor in a mammal, which method comprises administering to saidmammal an effective amount of a compound of the formula according toclaim
 1. 14. The method according to claim 13 wherein the mammal isafflicted with a chemokine mediated disease selected from the groupconsisting of psoriasis, atopic dermatitis, osteo arthritis, rheumatoidarthritis, asthma, chronic obstructive pulmonary disease, adultrespiratory distress syndrome, inflammatory bowel disease, Crohn'sdisease, ulcerative colitis, stroke, septic shock, multiple sclerosis,endotoxic shock, gram negative sepsis, toxic shock syndrome, cardiac andrenal reperfusion injury, glomerulonephritis, thrombosis, graft vs. hostreaction, alzheimers disease, allograft rejections, malaria, restenosis,angiogenesis, atherosclerosis, osteoporosis, gingivitis and undesiredhematopoietic stem cells release and diseases caused by respiratoryviruses, herpesviruses, and hepatitis viruses, meningitis, cysticfibrosis, pre-term labor, cough, pruritus, multi-organ dysfucntions,trauma, strains, sprains, contusions, psoriatic arthritis, herpes,encephalitis, CNS vasculitis, traumatic brain injury, CNS tumors,subarachnoid hemorrhage, post surgical trauma, interstitial pneumonitis,hypersensitivity, crystal induced arthritis, acute and chronicpancreatitis, acute alcoholic hepatitis, necrotizing enterocolitis,chronic sinusitis, uveitis, polymyositis, vasculitis, acne, gastric andduodenal ulcers, celiac disease, esophagitis, glossitis, airflowobstruction, airway hyperresponsiveness, bronchiolitis obliteransorganizing pneumonia, bronchiectasis, bronchiolitis, bronchiolitisobliterans, chronic bronchitis, cor pulmonae, dyspnea, emphysema,hypercapnea, hyperinflation, hypoxemia, hyperoxia-induced inflammations,hypoxia, surgerical lung volume reduction, pulmonary fibrosis, pulmonaryhypertension, right ventricular hypertropy, sarcoidosis, small airwaydisease, ventilation-perfusion mismatching, wheeze, colds and lupus.